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Friday, September 28, 2007

Transcription of the 7SL Gene

There are five different kinds of RNA polymerase in eukaryotes. Each of them is responsible for transcription of a different class of gene [Eukaryotic RNA Polymerases]. RNA polymerase III transcribes a heterogeneous class of genes that give rise to small RNAs.

The class III genes can be subdivided into four types depending on the location of the promoter regions. Type 1 genes possess an internal control region (ICR) that functions as the promoter. What this means is that the site of binding of the pol III transcription complex is within the gene. 5S RNA genes are the only kind of type 1 gene.

A cartoon drawing of the pol III transcription complex on a 5S RNA gene is shown above (Moran, Scrimgeour et al. 1994). The transcription factor TFIIIA binds to the internal control region (ICR). Another transcription factor, TFIIIC, binds TFIIIA and it, in turn, interacts with TFIIIB and RNA polymerase III. Transcription is initiated at a site (+1) upstream from the internal control region. Note that when the 5S RNA is produced it will contain the binding sites for TFIIIA. The significance of this fact will become clear in a few minutes.

The type 2 genes have an intragenic promoter with two binding sites, A and B. TFIIIC binds directly to this promoter causing the assembly of a transcription complex upstream in the same manner as the type 1 genes. Most of the transfer RNA (tRNA) genes are type 2 genes.

Type 3 genes have an upstream promoter and no internal promoter. In this sense they resemble the typical class II genes (transcribed by RNA polymerase II), such as those that encode protein. The U6 snRNA gene, a component of the spliceosome [RNA Splicing: Introns and Exons] is the prototype gene of this category.

Finally, type 4 genes have both an internal region where the regular class III transcription factors bind and a promoter at the 5′ end of the gene where regulatory transcription factors bind to control transcription. The 7SL RNA gene is a type 4 gene.

Recall that 7SL RNA is the major component of Signal Recognition Particle (SRP). There are three genes for this RNA located on human chromosome 14 [Human Genes Involved in the Signal Hypothesis Pathway]. Since 7SL RNA is one of the small RNAs present in most cells it should not come as a surprise that its gene is transcribed by RNA polymerase III.

The RN7SL promoter has been characterized by Englert et al. (2004). The main features are shown in the diagram above. The solid blue box represents the gene—the DNA sequence corresponding to the 7SL RNA. There's a start site for transcription (+1) that's determined by the positioning of RNA polymerase III upstream. The transcription complex is assembled when TFIIIC binds to an internal control region specified by box A and box B. These are short DNA sequences (10 bp) that resemble the binding sites in tRNA genes. The upstream promoter region consists of a TATA box where RNA polymerase III binds and a regulatory site where unknown transcription factors (TF) bind.

Transcription terminates at a short stretch of thymidylate residues (T) at the 3′ end of the gene.

All four regulatory sites have to be present for maximum rates of transcription but—and this is important—there will still be low levels of transcription if only the A box and the B box are present.

From time to time cellular RNAs are copied by an enzyme called reverse transcriptase to create a DNA:RNA double-stranded molecule. On occasion this hybrid molecule will become integrated into the genome through a nonhomologous recombination event. (Sometimes the RNA strand will be replaced by DNA synthesis to create double-stranded DNA corresponding to the 7SL sequence.) This creates something called a processed pseudogene. Most genomes have hundreds of these pseudogenes derived from hundreds of different genes. They have arisen from accidental events over the course of millions of year of evolution and since there's no pressure to eliminate them, they are retained in the genome as junk.

Junk DNA
If the processed pseudogene is derived from a class III gene there's a good chance that it will retain prompter activity because a good part of the promoter sequence was present in the RNA molecule. This is the case with 7SL pseudogenes and tRNA pseudogenes. They are often transcribed at low levels. The production of additional RNAs from the processed pseudogenes increases the probability that more pseudogenes will be created. More significantly, if the processed 7SL or tRNA pseudogenes happen to integrate near certain mobilization sequences they will be converted to retrotransposons because they can direct transcription of themselves—a necessary step in transposition. When this happens the pseudogenes will spread rapidly throughout the genome. We call this selfish DNA.

More than 10% of your genome consists of degenerate 7SL genes. That's where some of the junk DNA in your genome comes from.

Englert, M., Felis, M., Junker, V. and Beier, H. (2004) Novel upstream and intragenic control elements for the RNA polymerase III-dependent transcription of human 7SL RNA genes. Biochimie. 86:867-74. [PubMed]


Timothy V Reeves said...

I've read intoductory texts on genetics and transcription; it is clearly a lot more complicated than the 'cartoons' of 'cartoons' that I am familiar with! The more you 'zoom in' the more detail is revealed.

Anonymous said...

I can appreciate how far we've come from the Macintosh MacPaint, Pac-Man graphics of the '80s to depicting RNApol like the section of an intestine or perhaps a grub laying blue eggs.

Anonymous said...

Yes, now they look like balloon animals ;)