Maf1p, a negative effector of RNA polymerase III in Saccharomyces cerevisiae.
Although yeast RNA polymerase III (Pol III) and the auxiliary factors TFIIIC and TFIIIB are well characterized, the mechanisms of class III gene regulation are poorly understood. Previous studies identified MAF1, a gene that affects tRNA suppressor efficiency and interacts genetically with Pol III. We show here that tRNA levels are elevated in maf1 mutant cells. In keeping with the higher levels of tRNA observed in vivo, the in vitro rate of Pol III RNA synthesis is significantly increased in maf1 cell extracts. Mutations in the RPC160 gene encoding the largest subunit of Pol III which reduce tRNA levels were identified as suppressors of the maf1 growth defect. Interestingly, Maf1p is located in the nucleus and coimmunopurifies with epitope-tagged RNA Pol III. These results indicate that Maf1p acts as a negative effector of Pol III synthesis. This potential regulator of Pol III transcription is likely conserved since orthologs of Maf1p are present in other eukaryotes, including humans.
Pluta
K
K
Department of Genetics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, 02 106 Warsaw, Poland.
Lefebvre
O
O
Martin
N C
NC
Smagowicz
W J
WJ
Stanford
D R
DR
Ellis
S R
SR
Hopper
A K
AK
Sentenac
A
A
Boguta
M
M
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D002467:Cell Nucleus / Q000378:metabolism; D000939:Epitopes; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D015151:Immunoblotting; D008856:Microscopy, Fluorescence; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D012313:RNA / Q000378:metabolism; D012320:RNA Polymerase III / Q000737:chemistry / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D035361:Transcription Factor TFIIIB; D014157:Transcription Factors / Q000737:chemistry; D019474:Transcription Factors, TFIII / Q000737:chemistry; D014158:Transcription, Genetic; D000098832:Transcription Factor TFIIIC
D000939:Epitopes; D005656:Fungal Proteins; C104954:MAF1 protein, S cerevisiae; D012335:RNA, Ribosomal; D029701:Saccharomyces cerevisiae Proteins; D035361:Transcription Factor TFIIIB; D014157:Transcription Factors; D019474:Transcription Factors, TFIII; D000098832:Transcription Factor TFIIIC; D012313:RNA; D012343:RNA, Transfer; D012320:RNA Polymerase III