The fission yeast Rpb4 subunit of RNA polymerase II plays a specialized role in cell separation.
RNA polymerase II is a complex of 12 subunits, Rpb1 to Rpb12, whose specific roles are only partly understood. Rpb4 is essential in mammals and fission yeast, but not in budding yeast. To learn more about the roles of Rpb4, we expressed the rpb4 gene under the control of regulatable promoters of different strength in fission yeast. We demonstrate that below a critical level of transcription, Rpb4 affects cellular growth proportional to its expression levels: cells expressing lower levels of rpb4 grew slower compared to cells expressing higher levels. Lowered rpb4 expression did not affect cell survival under several stress conditions, but it caused specific defects in cell separation similar to sep mutants. Microarray analysis revealed that lowered rpb4 expression causes a global reduction in gene expression, but the transcript levels of a distinct subset of genes were particularly responsive to changes in rpb4 expression. These genes show some overlap with those regulated by the Sep1-Ace2 transcriptional cascade required for cell separation. Most notably, the gene expression signature of cells with lowered rpb4 expression was highly similar to those of mcs6, pmh1, sep10 and sep15 mutants. Mcs6 and Pmh1 encode orthologs of metazoan TFIIH-associated cyclin-dependent kinase (CDK)-activating kinase (Cdk7-cyclin H-Mat1), while Sep10 and Sep15 encode mediator components. Our results suggest that Rpb4, along with some other general transcription factors, plays a specialized role in a transcriptional pathway that controls the cell cycle-regulated transcription of a specific subset of genes involved in cell division.
Sharma
Nimisha
N
University School of Biotechnology, G.G.S. Indraprastha University, Kashmere Gate, Delhi, 110006, India.
Marguerat
Samuel
S
Mehta
Surbhi
S
Watt
Stephen
S
Bähler
Jürg
J
D002455:Cell Division / Q000235:genetics* / Q000502:physiology; D015966:Gene Expression Regulation, Fungal / Q000235:genetics*; D046228:Microarray Analysis; D008856:Microscopy, Fluorescence; D012319:RNA Polymerase II / Q000235:genetics* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012568:Schizosaccharomyces / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics* / Q000378:metabolism*
D029702:Schizosaccharomyces pombe Proteins; D012319:RNA Polymerase II; C499659:rpb4 protein, S pombe