Shapiro doubles down on his claim that junk DNA doesn't exist.
It's been a while since we've heard from James Shaprio. You might recall that James A. Shapiro is a biochemistry/microbiology professor at the University of Chicago and the author of a book promoting natural genetic engineering. I reviewed his book and didn't like it very much—Shapiro didn't like my review [James Shapiro Never Learns] [James Shapiro Responds to My Review of His Book].
Gert Korthof thinks that the current view of evolution is incomplete and he's looking for a better explanation. He just finished reading my book so he wrote a review on his blog.
Scientists say: 90% of your genome is junk. Have a nice day! Biochemist Laurence Moran defends junk DNA theory
The good news is that I've succeeded in making Gert Korthof think more seriously about junk DNA and random genetic drift. The bad news is that I seem to have given him the impression that natural selection is not an important part of evolution. Furthermore, he insists that "evolution needs both mutation and natural selection" because he doesn't like the idea that random genetic drift may be the most common mechanism of evolution. He thinks that statement only applies at the molecular level. But "evolution" doesn't just refer to adaptation at the level of organisms. It's just not true that all examples of evolution must involve natural selection.
I think I've failed to explain the null hypothesis correctly because Korthof writes,
It's clear this is a polemical book. It is a very forceful criticism of ENCODE and everyone who uncritically accepts and spreads their views including Nature and Science. I agree that this criticism is necessary. However, there is a downside. Moran writes that the ENCODE research goals of documenting all transcripts in the human genome was a waste of money. Only a relatively small group of transcripts have a proven biological function ("only 1000 lncRNAs out of 60,000 were conserved in mammals"; "the number with a proven function is less than 500 in humans"; "The correct null hypothesis is that these long noncoding RNAs are examples of noisy transcription", or junk RNA"). Furthermore, Moran also thinks it is a waste of time and money to identify the functions of the thousands of transcripts that have been found because he knows its all junk. I disagree. The null hypothesis is an hypothesis, not a fact. One cannot assume it is true. That would be the 'null dogma'.
That's a pretty serious misunderstanding of what I meant to say. I think it was a worthwhile effort to document the number of transcripts in various cell types and all the potential regulatory sequences. What I objected to was the assumption by ENCODE researchers that these transcripts and sites were functional simply because they exist. The null hypothesis is no function and scientists must provide evidence of function in order to refute the null hypothesis.
I think it would be a very good idea to stop further genomic surveys and start identifying which transcripts and putative regulatory elements are actually functional. I'd love to know the answer to that very important question. However, I recognize that it will be expensive and time consuming to investigate every transcript and every putative regulatory element. I don't think any lab is going to assign random transcripts and random transcription factor binding sites to graduate students and postdocs because I suspect that most of those sequences aren't going to have a function. If I were giving out grant money I give it to some other lab. In that sense, I believe that it would be a waste of time and money to search for the function of tens of thousands of transcripts and over one million transcription factor binding sites.
That not dogmatic. It's common sense. Most of those transcripts and binding sites are not conserved and not under purifying election. That's pretty good evidence that they aren't functional, especially if you believe in the importance of natural selection.
There's lot more to his review including some interesting appendices. I recommend that you read it carefully to see a different perspective than the one I adocate in my book.
Francis Collins, and many others, believe that the concept of junk DNA is outmoded because recent discoveries have shown that most of the human genome is devoted to regulation. This is part of a clash of worldviews where one side sees the genome as analogous to a finely tuned Swiss watch with no room for junk and the other sees the genome as a sloppy entity that's just good enough to survive.
The ENCODE researchers and their allies claim that the human genome contains more than 600,000 regulatory sites and that means an average of 24 per gene covering about 10,000 bp per gene. I explain why these numbers are unreasonable and why most of the sites they identify have nothing to do with biologically significant regulation.
This chapter also covers the epigenetics hype and restriction/modification.
Click on this link to see more. Chapter 10: Turning Genes On and Off
In September 2012, the ENCODE researchers published a bunch of papers claiming to show that 80% of the human genome was functional. They helped orchestrate a massive publicity campaign with the help pf Nature— a campaign that succeeded in spreading the message that junk DNA had been refuted.
That claim was challenged within 24 hours by numerous scientists on social media. They pointed out that the ENCODE researchers were using a ridiculous definition of function and that they had completely ignored all the evidence for junk DNA. Over the next two years there were numerous scientific papers criticizing the ENCODE claims and the ENCODE researchers were forced to retract the claim that they had proven that 80% of the genome is functional.
I discuss what went wrong and lay the blame mostly on the ENCODE researchers who did not behave as proper scientists when presenting a controversial hypothesis. The editors of Nature share the blame for not doing a proper job of vetting the ENCODE claims and not subjecting the papers to rigorous peer review. Science writers also failed to think critically about the results they were reporting.
Click on this link to see more. Chapter 9: The ENCODE Publicity Campaign
If you are interested in learning more about junk DNA here's some links to relevant information.
Prologue Chapter 1: Introducing Genomes Chapter 2: The Evolution of Sloppy Genomes Chapter 3: Repetitive DNA and Mobile Genetic Elements Chapter 4: Why Don't Mutations Kill Us? Chaper 5: The Big Picture Chapter 6: How Many Genes? How Many Proteins? Chapter 7: Gene Families and the Birth and Death of Genes Chapter 8: Noncoding Genes and Junk RNA Chapter 9: The ENCODE Publicity Campaign Chapter 10: Turning Genes On and Off Chapter 11: Zen and the Art of Coping with a Sloppy Genome
The correct definition of a molecular gene isn't difficult but getting it recognized and accepted is a different story.
When writing my book on junk DNA I realized that there was an issue with genes. The average scientist, and consequently the average science writer, has a very confused picture of genes and the proper way to define them. The issue shouldn't be confusing for Sandwalk readers since we've covered that ground many times in the past. I think the best working definition of a gene is, "A gene is a DNA sequence that is transcribed to produce a functional product" [What Is a Gene?]
The 2022 Nobel Prize in Physiology or Medicne went to Svante Pääbo “for his discoveries concerning the genomes of extinct hominins and human evolution”. It's one of a long list of Nobel Prizes awarded for technological achievement. It most cases, the new techniques led to a better understanding of science and medicine.
Since World War II, there have been significant advances in our understanding of biology but most of these have come about by the slow and steady accumulation of knowledge and not by paradigm-shifting breakthroughs. These advances don't often get recognized by the Nobel Prize committees because it's difficult to single out any one individual or any single experiment that merits a Nobel Prize. In some cases the Nobel Prize committees have tried to recognize major advances by picking out leaders that have made important contributions over a number of years but their choices don't always satisfy others in the field. One of the notable successes is the awarding of Nobel Prizes to Max Delbrück, Alfred D. Hershey and Salvador E. Luria “for their discoveries concerning the replication mechanism and the genetic structure of viruses” (Nobel Prize in Physiology or Medicine 1969). Another is Edward B. Lewis, Christiane Nüsslein-Volhard and Eric F. Wieschaus “for their discoveries concerning the genetic control of early embryonic development” (Nobel Prize in Physiology or Medicine 1995)
There's an old saying that birds of a feather flock together. It means that people with the same interests tend to associate with each other. It's extended meaning refers to the fact that people who believe in one thing (X) tend to also believe in another (Y). It usually means that X and Y are both questionable beliefs and it's not clear why they should be associated.
I've noticed an association between those who promote epigenetics far beyond it's reasonable limits and those who reject junk DNA in favor of a genome that's mostly functional. There's no obvious reason why these two beliefs should be associated with each other but they are. I assume it's related to the idea that both beliefs are presumed to be radical departures from the standard dogma so they reinforce the idea that the author is a revolutionary.
Or maybe it's just that sloppy thinking in one field means that sloppy thinking is the common thread.
Here's an example from Chapter 4 of a 2023 edition of the Handbook of Epigenetics (Third Edition).
The central dogma of life had clearly established the importance of the RNA molecule in the flow of genetic information. The understanding of transcription and translation processes further elucidated three distinct classes of RNA: mRNA, tRNA and rRNA. mRNA carries the information from DNA and gets translated to structural or functional proteins; hence, they are referred to as the coding RNA (RNA which codes for proteins). tRNA and rRNA help in the process of translation among other functions. A major part of the DNA, however, does not code for proteins and was previously referred to as junk DNA. The scientists started realizing the role of the junk DNA in the late 1990s and the ENCODE project, initiated in 2003, proved the significance of junk DNA beyond any doubt. Many RNA types are now known to be transcribed from DNA in the same way as mRNA, but unlike mRNA they do not get translated into any protein; hence, they are collectively referred to as noncoding RNA (ncRNA). The studies have revealed that up to 90% of the eukaryotic genome is transcribed but only 1%–2% of these transcripts code for proteins, the rest all are ncRNAs. The ncRNAs less than 200 nucleotides are called small noncoding RNAs and greater than 200 nucleotides are called long noncoding RNAs (lncRNAs).
In case you haven't been following my blog posts for the past 17 years, allow me to briefly summarize the flaws in that paragraph.
So, I ask the same question that I've been asking for decades. How does this stuff get published?
In order to have a productive discussion about junk DNA we needed to agree on how to define "function" and "junk." Disagreements over the definitions spawned the Function Wars that became intense over the past decade. That war is over and now it's time to move beyond nitpicking about terminology.
The idea that most of the human genome is composed of junk DNA arose gradually in the late 1960s and early 1970s. The concept was based on a lot of evidence dating back to the 1940s and it gained support with the discovery of massive amounts of repetitive DNA.
Various classes of functional DNA were known back then including: regulatory sequences, protein-coding genes, noncoding genes, centromeres, and origins of replication. Other categories have been added since then but the total amount of functional DNA was not thought to be more than 10% of the genome. This was confirmed with the publication of the human genome sequence.
From the very beginning, the distinction between functional DNA and junk DNA was based on evolutionary principles. Functional DNA was the product of natural selection and junk DNA was not constrained by selection. The genetic load argument was a key feature of Susumu Ohno's conclusion that 90% of our genome is junk (Ohno, 1972a; Ohno, 1972b).
Most of our genome is transcribed but so far only a small number of these transcripts have a well-established biological function.
The fact that most of our genome is transcribed has been known for 50 years but that fact only became widely known with the publication of ENCODE's preliminary results in 2007 (ENCODE, 2007). The ENOCDE scientists referred to this as "pervasive transription" and this label has stuck.
By the end of the 1970s we knew that much of this transcription was due to introns. The latest data shows that protein coding genes and known noncoding genes occupy about 45% of the genome and most of that is intron sequences that are mostly junk. That leaves 30-40% of the genome that is transcribed at some point producing something like one million transcripts of unknown function.
A new master's thesis on the history of junk DNA has been posted. It's from the Department of History at the University of Chicago.
My routine scan for articles on junk DNA turned up the abstract of an M.A. thesis on the history of junk DNA: Requiem for a Gene: The Problem of Junk DNA for the Molecular Paradigm. The supervisor is Professor Emily Kern in the Department of History at the University of Chicago. I've written to her to ask for a copy of the thesis and for permission to ask her, and her student, some questions about the thesis. No reply so far.
Here's the abstract of the thesis.
“Junk DNA” has been at the center of several high-profile scientific controversies over the past four decades, most recently in the disputes over the ENCODE Project. Despite its prominence in these debates, the concept has yet to be properly historicized. In this thesis, I seek to redress this oversight, inaugurating the study of junk DNA as a historical object and establishing the need for an earlier genesis for the concept than scholars have previously recognized. In search of a new origin story for junk, I chronicle developments in the recognition and characterization of noncoding DNA sequences, positioning them within existing historiographical narratives. Ultimately, I trace the origin of junk to 1958, when a series of unexpected findings in bacteria revealed the existence of significant stretches of DNA that did not encode protein. I show that the discovery of noncoding DNA sequences undermined molecular biologists’ vision of a gene as a line of one-dimensional code and, in turn, provoked the first major crisis in their nascent field. It is from this crisis, I argue, that the concept of junk DNA emerged. Moreover, I challenge the received narrative of junk DNA as an uncritical reification of the burgeoning molecular paradigm. By separating the history of junk DNA from its mythology, I demonstrate that the conceptualization of junk DNA reveals not the strength of molecular biological authority but its fragility.
It looks like it might be a history of noncoding DNA but I won't know for certain until I see the entire thesis. It's only available to students and staff at the University of Chicago.
There's a lot of talk about ChatGPT and how it can prepare lectures and get good grades on undergraduate exams. However, ChatGPT is only as good as the information that's popular on the internet and that's not always enough to get a good grade on my exam.
ChatGPT is an artificial intelligence (AI) program that's designed to answer questions using a style and language that's very much like the responses you would get from a real person. It was developed by OpenAI, a tech company in San Francisco. You can create an account and log in to ask any question you want.
Several professors have challenged it with exam questions and they report that ChatGPT would easily pass their exams. I was skeptical, especially when it came to answering questions on controversial topics where there was no clear answer. I also suspected that ChatGPT would get it's answers from the internet and this means that popular, but incorrect, views would likely be part of ChatGPT's response.
Here are my questions and the AI program's answers. It did quite well in some cases but not so well in others. My main concern is that programs like this might be judged to be reliable sources of information despite the fact that the real source is suspect.
In spite of what you might have read, the human genome does not contain one million functional enhancers.
The Sept. 15, 2022 issue of Nature contains a news article on "Gene regulation" [Two-layer design protects genes from mutations in their enhancers]. It begins with the following sentence.
The human genome contains only about 20,000 protein-coding genes, yet gene expression is controlled by around one million regulatory DNA elements called enhancers.
Sandwalk readers won't need to be told the reference for such an outlandish claim because you all know that it's the ENCODE Consortium summary paper from 2012—the one that kicked off their publicity campaign to convince everyone of the death of junk DNA (ENCODE, 2012). ENCODE identified several hundred thousand transcription factor (TF) binding sites and in 2012 they estimated that the total number of base pairs invovled in regulating gene expression could account for 20% of the genome.
How many of those transcription factor binding sites are functional and how many are due to spurious binding to sites that have nothing to do with gene regulation? We don't know the answer to that question but we do know that there will be a huge number of spurious binding sites in a genome of more than three billion base pairs [Are most transcription factor binding sites functional?].
The scientists in the ENCODE Consortium didn't know the answer either but what's surprising is that they didn't even know there was a question. It never occured to them that some of those transcription factor binding sites have nothng to do with regulation.
Fast forward ten years to 2022. Dozens of papers have been published criticizing the ENCODE Consortium for their stupidity lack of knowledge of the basic biochemical properties of DNA binding proteins. Surely nobody who is interested in this topic believes that there are one million functional regulatory elements (enhancers) in the human genome?
Wrong! The authors of this Nature article, Ran Elkon at Tel Aviv University (Israel) and Reuven Agami at the Netherlands Cancer Institute (Amsterdam, Netherlands), didn't get the message. They think it's quite plausible that the expression of every human protein-coding gene is controlled by an average of 50 regulatory sites even though there's not a single known example any such gene.
Not only that, for some reason they think it's only important to mention protein-coding genes in spite of the fact that the reference they give for 20,000 protein-coding genes (Nurk et al., 2022) also claims there are an additional 40,000 noncoding genes. This is an incorrect claim since Nurk et al. have no proof that all those transcribed regions are actually genes but let's play along and assume that there really are 60,000 genes in the human genome. That reduces the average number of enhancers to an average of "only" 17 enhancers per gene. I don't know of a single gene that has 17 or more proven enhancers, do you?
Why would two researchers who study gene regulation say that the human genome contains one million enhancers when there's no evidence to support such a claim and it doesn't make any sense? Why would Nature publish this paper when surely the editors must be aware of all the criticism that arose out of the 2012 ENCODE publicity fiasco?
I can think of only two answers to the first question. Either Elkon and Agami don't know of any papers challenging the view that most TF binding sites are functional (see below) or they do know of those papers but choose to ignore them. Neither answer is acceptable.
I think that the most important question in human gene regulation is how much of the genome is devoted to regulation. How many potential regulatory sites (enhancers) are functional and how many are spurious non-functional sites? Any paper on regulation that does not mention this problem should not be published. All results have to interpreted in light of conflicting claims about function.
Here are some example of papers that raise the issue. The point is not to prove that these authors are correct - although they are correct - but to show that there's a controvesy. You can't just state that there are one million regulatory sites as if it were a fact when you know that the results are being challenged.
"The observations in the ENCODE articles can be explained by the fact that biological systems are noisy: transcription factors can interact at many nonfunctional sites, and transcription initiation takes place at different positions corresponding to sequences similar to promoter sequences, simply because biological systems are not tightly controlled." (Morange, 2014)"... ENCODE had not shown what fraction of these activities play any substantive role in gene regulation, nor was the project designed to show that. There are other well-studied explanations for reproducible biochemical activities besides crucial human gene regulation, including residual activities (pseudogenes), functions in the molecular features that infest eukaryotic genomes (transposons, viruses, and other mobile elements), and noise." (Eddy, 2013)
"Given that experiments performed in a diverse number of eukaryotic systems have found only a small correlation between TF-binding events and mRNA expression, it appears that in most cases only a fraction of TF-binding sites significantly impacts local gene expression." (Palazzo and Gregory, 2014)
One surprising finding from the early genome-wide ChIP studies was that TF binding is widespread, with thousand to tens of thousands of binding events for many TFs. These number do not fit with existing ideas of the regulatory network structure, in which TFs were generally expected to regulate a few hundred genes, at most. Binding is not necessarily equivalent to regulation, and it is likely that only a small fraction of all binding events will have an important impact on gene expression. (Slattery et al., 2014)
Detailed maps of transcription factor (TF)-bound genomic regions are being produced by consortium-driven efforts such as ENCODE, yet the sequence features that distinguish functional cis-regulatory sites from the millions of spurious motif occurrences in large eukaryotic genomes are poorly understood. (White et al., 2013)
One outstanding issue is the fraction of factor binding in the genome that is "functional", which we define here to mean that disturbing the protein-DNA interaction leads to a measurable downstream effect on gene regulation. (Cusanovich et al., 2014)
... we expect, for example, accidental transcription factor-DNA binding to go on at some rate, so assuming that transcription equals function is not good enough. The null hypothesis after all is that most transcription is spurious and alterantive transcripts are a consequence of error-prone splicing. (Hurst, 2013)
... as a chemist, let me say that I don't find the binding of DNA-binding proteins to random, non-functional stretches of DNA surprising at all. That hardly makes these stretches physiologically important. If evolution is messy, chemistry is equally messy. Molecules stick to many other molecules, and not every one of these interactions has to lead to a physiological event. DNA-binding proteins that are designed to bind to specific DNA sequences would be expected to have some affinity for non-specific sequences just by chance; a negatively charged group could interact with a positively charged one, an aromatic ring could insert between DNA base pairs and a greasy side chain might nestle into a pocket by displacing water molecules. It was a pity the authors of ENCODE decided to define biological functionality partly in terms of chemical interactions which may or may not be biologically relevant. (Jogalekar, 2012)
Nurk, S., Koren, S., Rhie, A., Rautiainen, M., Bzikadze, A. V., Mikheenko, A., et al. (2022) The complete sequence of a human genome. Science, 376:44-53. [doi:10.1126/science.abj6987]
The ENCODE Project Consortium (2012) An integrated encyclopedia of DNA elements in the human genome. Nature, 489:57-74. [doi: 10.1038/nature11247]
Here's an example of a press release that distorts science by promoting incorrect information that is not found in the actual publication.
The problems with press releases are well-known but nobody is doing anything about it. I really like the discussion in Stuart Ritchie's recent (2020) book where he begins with the famous "arsenic affair" in 2010. Sandwalk readers will recall that this started with a press conference by NASA announcing that arsenic replaces phosphorus in the DNA of some bacteria. The announcement was treated with contempt by the blogosphere and eventually the claim was discproved by Rosie Redfield who showed that the experiment was flawed [The Arsenic Affair: No Arsenic in DNA!].
This was a case where the science was wrong and NASA should have known before it called a press conference. Ritchie goes on to document many cases where press releases have distorted the science in the actual publication. He doesn't mention the most egregious example, the ENCODE publicity campaign that successfully convinced most scientists that junk DNA was dead [The 10th anniversary of the ENCODE publicity campaign fiasco].
I like what he says about "churnalism" ...
In an age of 'churnalism', where time-pressed journalists often simply repeat the content of press releases in their articles (science news reports are often worded vitrually identically to a press release), scientists have a great deal of power—and a great deal of responsibility. The constraints of peer review, lax as they might be, aren't present at all when engaging with the media, and scientists' biases about the importance of their results can emerge unchecked. Frustratingly, once the hype bubble has been inflated by a press release, it's difficult to burst.
Press releases of all sorts are failing us but university press releases are the most disappointing because we expect universities to be credible sources of information. It's obvious that scientists have to accept the blame for deliberately distorting their findings but surely the information offices at universities are also at fault? I once suggested that every press release has to include a statement, signed by the scientists, saying that the press release accurately reports the results and conclusions that are in the published article and does not contain any additional information or speculation that has not passed peer review.
Let's look at a recent example where the scientists would not have been able to truthfully sign such a statement.
A group of scientists based largely at The University of Sheffield in Sheffield (UK) recently published a paper in Nature on DNA damage in the human genome. They noted that such damage occurs preferentially at promoters and enhancers and is associated with demethylation and transcription activation. They presented evidence that the genome can be partially protected by a protein called "NuMA." I'll show you the abstract below but for now that's all you need to know.
The University of Sheffield decided to promote itself by issuing a press release: Breaks in ‘junk’ DNA give scientists new insight into neurological disorders. This title is a bit of a surprise since the paper only talks about breaks in enhancers and promoters and the word "junk" doesn't appear anywhere in the published report in Nature.
The first paragraph of the press release isn' very helpful.
‘Junk’ DNA could unlock new treatments for neurological disorders as scientists discover how its breaks and repairs affect our protection against neurological disease.
What could this mean? Surely they don't mean to imply that enhancers and promoters are "junk DNA"? That would be really, really, stupid. The rest of the press release should explain what they mean.
The groundbreaking research from the University of Sheffield’s Neuroscience Institute and Healthy Lifespan Institute gives important new insights into so-called junk DNA—or DNA previously thought to be non-essential to the coding of our genome—and how it impacts on neurological disorders such as Motor Neurone Disease (MND) and Alzheimer’s.
Until now, the body’s repair of junk DNA, which can make up 98 per cent of DNA, has been largely overlooked by scientists, but the new study published in Nature found it is much more vulnerable to breaks from oxidative genomic damage than previously thought. This has vital implications on the development of neurological disorders.
Oops! Apparently, they really are that stupid. The scientists who did this work seem to think that 98% of our genome is junk and that includes all the regulatory sequences. It seems like they are completely unaware of decades of work on discovering the function of these regulatory sequences. According The University of Sheffield, these regulatory sequences have been "largely overlooked by scientists." That will come as a big surprise to many of my colleagues who worked on gene regulation in the 1980s and in all the decades since then. It will probably also be a surprise to biochemistry and molecular biology undergraduates at Sheffield—at least I hope it will be a surprise.
Professor Sherif El-Khamisy, Chair in Molecular Medicine at the University of Sheffield, Co-founder and Deputy Director of the Healthy Lifespan Institute, said: “Until now the repair of what people thought is junk DNA has been mostly overlooked, but our study has shown it may have vital implications on the onset and progression of neurological disease."
I wonder if Professor Sherif El-Khamisy can name a single credible scientist who thinks that regulatory sequences are junk DNA?
There's no excuse for propagating this kind of misinformation about junk DNA. It's completely unnecessary and serves only to discredit the university and its scientists.
Ray, S., Abugable, A.A., Parker, J., Liversidge, K., Palminha, N.M., Liao, C., Acosta-Martin, A.E., Souza, C.D.S., Jurga, M., Sudbery, I. and El-Khamisy, S.F. (2022) A mechanism for oxidative damage repair at gene regulatory elements. Nature, 609:1038-1047. doi:[doi: 10.1038/s41586-022-05217-8]
Oxidative genome damage is an unavoidable consequence of cellular metabolism. It arises at gene regulatory elements by epigenetic demethylation during transcriptional activation1,2. Here we show that promoters are protected from oxidative damage via a process mediated by the nuclear mitotic apparatus protein NuMA (also known as NUMA1). NuMA exhibits genomic occupancy approximately 100 bp around transcription start sites. It binds the initiating form of RNA polymerase II, pause-release factors and single-strand break repair (SSBR) components such as TDP1. The binding is increased on chromatin following oxidative damage, and TDP1 enrichment at damaged chromatin is facilitated by NuMA. Depletion of NuMA increases oxidative damage at promoters. NuMA promotes transcription by limiting the polyADP-ribosylation of RNA polymerase II, increasing its availability and release from pausing at promoters. Metabolic labelling of nascent RNA identifies genes that depend on NuMA for transcription including immediate–early response genes. Complementation of NuMA-deficient cells with a mutant that mediates binding to SSBR, or a mitotic separation-of-function mutant, restores SSBR defects. These findings underscore the importance of oxidative DNA damage repair at gene regulatory elements and describe a process that fulfils this function.
On Sept. 5, 2012 ENCODE researchers, in collaboration with the science journal Nature, launched a massive publicity campaign to convince the world that junk DNA was dead. We are still dealing with the fallout from that disaster.
The Encyclopedia of DNA Elements (ENCODE) was originally set up to discover all of the functional elements in the human genome. They carried out a massive number of experiments involving a huge group of researchers from many different countries. The results of this work were published in a series of papers in the September 6th, 2012 issue of Nature. (The papers appeared on Sept. 5th.)
The ENCODE article on Wikipedia is a pretty good example of how to write a science article. Unfortunately, there are a few issues that will be very difficult to fix.
When Wikipedia was formed twenty years ago, there were many people who were skeptical about the concept of a free crowdsourced encyclopedia. Most people understood that a reliable source of information was needed for the internet because the traditional encyclopedias were too expensive, but could it be done by relying on volunteers to write articles that could be trusted?
The answer is mostly “yes” although that comes with some qualifications. Many science articles are not good; they contain inaccurate and misleading information and often don’t represent the scientific consensus. They also tend to be disjointed and unreadable. On the other hand, many non-science articles are at least as good, and often better, than anything in the traditional encyclopedias (eg. Battle of Waterloo; Toronto, Ontario; The Beach Boys).
By 2008, Wikipedia had expanded enormously and the quality of articles was being compared favorably to those of Encyclopedia Britannica, which had been forced to go online to compete. However, this comparison is a bit unfair since it downplays science articles.
ENCODE has mostly abandoned it's definition of function based on biochemical activity and replaced it with "candidate" function or "likely" function, but the message isn't getting out.
Back in 2012, the ENCODE Consortium announced that 80% of the human genome was functional and junk DNA was dead [What did the ENCODE Consortium say in 2012?]. This claim was widely disputed, causing the ENCODE Consortium leaders to back down in 2014 and restate their goal (Kellis et al. 2014). The new goal is merely to map all the potential functional elements.
... the Encyclopedia of DNA Elements Project [ENCODE] was launched to contribute maps of RNA transcripts, transcriptional regulator binding sites, and chromatin states in many cell types.
The new goal was repeated when the ENCODE III results were published in 2020, although you had to read carefully to recognize that they were no longer claiming to identify functional elements in the genome and they were raising no objections to junk DNA [ENCODE 3: A lesson in obfuscation and opaqueness].
The Wikipedia article on the Human genome contained a reference that I had not seen before.
"Finally DNA that is deleterious to the organism and is under negative selective pressure is called garbage DNA.[43]"
Reference 43 is a chapter in a book.
Pena S.D. (2021) "An Overview of the Human Genome: Coding DNA and Non-Coding DNA". In Haddad LA (ed.). Human Genome Structure, Function and Clinical Considerations. Cham: Springer Nature. pp. 5–7. ISBN 978-3-03-073151-9.
Sérgio Danilo Junho Pena is a human geneticist and professor in the Dept. of Biochemistry and Immunology at the Federal University of Minas Gerais in Belo Horizonte, Brazil. He is a member of the Human Genome Organization council. If you click on the Wikipedia link, it takes you to an excerpt from the book where S.D.J. Pena discusses "Coding and Non-coding DNA."
There are two quotations from that chapter that caught my eye. The first one is,
"Less than 2% of the human genome corresponds to protein-coding genes. The functional role of the remaining 98%, apart from repetitive sequences (constitutive heterochromatin) that appear to have a structural role in the chromosome, is a matter of controversy. Evolutionary evidence suggests that this noncoding DNA has no function—hence the common name of 'junk DNA.'"
Professor Pena then goes on to discuss the ENCODE results pointing out that there are many scientists who disagree with the conclusion that 80% of our genome is functional. He then says,
"Many evolutionary biologists have stuck to their guns in defense of the traditional and evolutionary view that non-coding DNA is 'junk DNA.'"
This is immediately followed by a quote from Dan Graur, implying that he (Graur) is one of the evolutionary biologists who defend the evolutionary view that noncoding DNA is junk.
I'm very interested in tracking down the reason for equating noncoding DNA and junk DNA, especially in contexts where the claim is obviously wrong. So I wrote to Professor Pena—he got his Ph.D. in Canada—and asked him for a primary source that supports the claim that "evolutionary science suggests that this noncoding DNA has no function."
He was kind enough to reply saying that there are multiple sources and he sent me links to two of them. Here's the first one.
I explained that this was somewhat ironic since I had written most of the Wikipedia article on Non-coding DNA and my goal was to refute the idea than noncoding DNA and junk DNA were synonyms. I explained that under the section on 'junk DNA' he would see the following statement that I inserted after writing sections on all those functional noncoding DNA elements.
"Junk DNA is often confused with non-coding DNA[48] but, as documented above, there are substantial fractions of non-coding DNA that have well-defined functions such as regulation, non-coding genes, origins of replication, telomeres, centromeres, and chromatin organizing sites (SARs)."
That's intended to dispel the notion that proponents of junk DNA ever equated noncoding DNA and junk DNA. I suggested that he couldn't use that source as support for his statement.
Here's my response to his second source.
The second reference is to a 2007 article by Wojciech Makalowski,1 a prominent opponent of junk DNA. He says, "In 1972 the late geneticist Susumu Ohno coined the term "junk DNA" to describe all noncoding sections of a genome" but that is a demonstrably false statement in two respects.
First, Ohno did not coin the term "junk DNA" - it was commonly used in discussions about genomes and even appeared in print many years before Ohno's paper. Second, Ohno specifically addresses regulatory sequences in his paper so it's clear that he knew about functional noncoding DNA that was not junk. He also mentions centromeres and I think it's safe to assume that he knew about ribosomal RNA genes and tRNA genes.
The only possible conclusion is that Makalowski is wrong on two counts.
I then asked about the second statement in Professor Pena's article and suggested that it might have been much better to say, "Many evolutionary biologists have stuck to their guns and defend the view that most of human genome is junk." He agreed.
So, what have we learned? Professor Pena is a well-respected scientist and an expert on the human genome. He is on the council of the Human Genome Organization. Yet, he propagated the common myth that noncoding DNA is junk and saw nothing wrong with Makalowski's false reference to Susumu Ohno. Professor Pena himself must be well aware of functional noncoding elements such as regulatory sequences and noncoding genes so it's difficult explain why he would imagine that prominant defenders of junk DNA don't know this.
I think the explanation is that this connection between noncoding DNA and junk DNA is so entrenched in the popular and scientific literature that it is just repeated as a meme without ever considering whether it makes sense.
1. The pdf appears to be a response to a query in Scientific American on February 12, 2007. It may be connected to a Scientific American paper by Khajavinia and Makalowski (2007).Khajavinia, A., and Makalowski, W. (2007) What is" junk" DNA, and what is it worth? Scientific American, 296:104. [PubMed]
Just before getting banned from Wikipedia, I was about to deal with a claim on the Intergenic region article. I had already fixed most of the other problems but there is still this statement in the subsection labeled "Properties."
According to the ENCODE project's study of the human genome, due to "both the expansion of genic regions by the discovery of new isoforms and the identification of novel intergenic transcripts, there has been a marked increase in the number of intergenic regions (from 32,481 to 60,250) due to their fragmentation and a decrease in their lengths (from 14,170 bp to 3,949 bp median length)"[2]
The source is one of the ENCODE papers published in the September 6 edition of Nature (Djebali et al., 2012). The quotation is accurate. Here's the full quotation.
As a consequence of both the expansion of genic regions by the discovery of new isoforms and the identification of novel intergenic transcripts, there has been a marked increase in the number of intergenic regions (from 32,481 to 60,250) due to their fragmentation and a decrease in their lengths (from 14,170 bp to 3,949 bp median length.
What's interesting about that data is what it reveals about the percentage of the genome devoted to intergenic DNA and the percentage devoted to genes. The authors claim that there are 60,250 intergenic regions, which means that there must be more than 60,000 genes.1 The median length of these intergenic regions is 3,949 bp and that means that roughly 204.5 x 106 bp are found in intergenic DNA. That's roughly 7% of the genome depending on which genome size you use. It doesn't mean that all the rest is genes but it sounds like they're saying that about 90% of the genome is occupied by genes.
In case you doubt that's what they're saying, read the rest of the paragraph in the paper.
Concordantly, we observed an increased overlap of genic regions. As the determination of genic regions is currently defined by the cumulative lengths of the isoforms and their genetic association to phenotypic characteristics, the likely continued reduction in the lengths of intergenic regions will steadily lead to the overlap of most genes previously assumed to be distinct genetic loci. This supports and is consistent with earlier observations of a highly interleaved transcribed genome, but more importantly, prompts the reconsideration of the definition of a gene.
It sounds like they are anticipating a time when the discovery of more noncoding genes will eventually lead to a situation where the intergenic regions disappear and all genes will overlap.
Now, as most of you know, the ENCODE papers have been discredited and hardly any knowledgeable scientist thinks there are 60,000 genes that occupy 90% of the genome. But here's the problem. I probably couldn't delete that sentence from Wikipedia because it meets all the criteria of a reliable source (published in Nature by scientists from reputable universities). Recent experience tells me that the Wikipolice Wikipedia editors would have blocked me from deleting it.
The best I could do would be to balance the claim with one from another "reliable source" such as Piovasan et al. (2019) who list the total number of exons and introns and their average sizes allowing you to calculate that protein-coding genes occupy about 35% of the genome. Other papers give slightly higher values for protein-coding genes.
It's hard to get a reliable source on the real number of noncoding genes and their average size but I estimate that there are about 5,000 genes and a generous estimate that they could take up a few percent of the genome. I assume in my upcoming book that genes probably occupy about 45% of the genome because I'm trying to err on the side of function.
An article on Intergenic regions is not really the place to get into a discussion about the number of noncoding genes but in the absence of such a well-sourced explanation the audience will be left with the statement from Djebali et al. and that's extremely misleading. Thus, my preference would be to replace it with a link to some other article where the controversy can be explained, preferably a new article on junk DNA.2
I was going to say,
The total amount of intergenic DNA depends on the size of the genome, the number of genes, and the length of each gene. That can vary widely from species to species. The value for the human genome is controversial because there is no widespread agreement on the number of genes but it's almost certain that intergenic DNA takes up at least 40% of the genome.
I can't supply a specific reference for this statement so it would never have gotten past the Wikipolice Wikpipedia editors. This is a problem that can't be solved because any serious attempt to fix it will probably lead to getting blocked on Wikipedia.
There is one other statement in that section in the article on Intergenic region.
Scientists have now artificially synthesized proteins from intergenic regions.[3]
I would have removed that statement because it's irrelevant. It does not contribute to understanding intergenic regions. It's undoubtedly one of those little factoids that someone has stumbled across and thinks it needs to be on Wikipedia.
Deletion of a statement like that would have met with fierce resistance from the Wikipedia editors because it is properly sourced. The reference is to a 2009 paper in the Journal of Biological Engineering: "Synthesizing non-natural parts from natural genomic template."
1. There are no intergenic regions between the last genes on the end of a chromosome and the telomeres.
2. The Wikipedia editors deleted the Junk DNA article about ten years ago on the grounds that junk DNA had been disproven.
Djebali, S., Davis, C. A., Merkel, A., Dobin, A., Lassmann, T., Mortazavi, A. et al. (2012) Landscape of transcription in human cells. Nature 489:101-108. [doi: 10.1038/nature11233]
Piovesan, A., Antonaros, F., Vitale, L., Strippoli, P., Pelleri, M. C., and Caracausi, M. (2019) Human protein-coding genes and gene feature statistics in 2019. BMC research notes 12:315. [doi: 10.1186/s13104-019-4343-8]
Functional DNA is best defined as DNA that is currently under purifying selection. In other words, it can't be deleted without affecting the fitness of the individual. This is the "maintenance function" definition and it differs from the "causal role" and "selected effect" definitions [The Function Wars Part IX: Stefan Linquist on Causal Role vs Selected Effect].
It has always been difficult to determine whether a given sequence is under purifying selection so sequence conservation is often used as a proxy. This is perfectly justifiable since the two criteria are strongly correlated. As a general rule, sequences that are currently being maintained by selection are ancient enough to show evidence of conservation. The only exceptions are de novo sequences and sequences that have recently become expendable and these are rare.
