Although yeast RNA polymerase III (Pol III) and the auxiliary factors TFIII
C and TFIIIB are well characterized, the mechanisms of class III gene regul
ation 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. Mutatio
ns in the RPC160 gene encoding the largest subunit of Pol III which reduce
tRNA levels were identified as suppressors of the maf1 growth defect. Inter
estingly, Maf1p is located in the nucleus and coimmunopurifies with epitope
-tagged RNA Pol III. These results indicate that Maf1p acts as a negative e
ffector of Pol III synthesis. This potential regulator of Pol III transcrip
tion is likely conserved since orthologs of Maf1p are present in other euka
ryotes, including humans.