Maf1p, a Negative Effector of RNA Polymerase III in Saccharomyces cerevisiae

Abstract

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 maf1cell 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.

ACKNOWLEDGMENTS

We thank B. Szczesniak and M. Steffen for excellent technical assistance. We are also grateful to Joël Acker, Christine Conesa, Gérald Peyroche, and Emmanuel Favry (CEA/Saclay) for helpful discussions and their help with the in vitro experiments.

This work was supported by State Committee for Scientific Research (KBN) grant 6PO4B02915 to K.P. and M.B., State Committee for Scientific Research grant 6P04A03315 to K.P. National Science Foundation grant MCB 9506810 to A.K.H., National Science Foundation grant MCB9528216 to N.C.M., and French-Polish Centre of Biotechnology of Plants grants to K.P. and W.J.S.