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Genetics, Vol 143, 1149-1161, Copyright © 1996
INVESTIGATIONS |
RNA Polymerase III Defects Suppress a Conditional-Lethal Poly(A) Polymerase Mutation in Saccharomyces cerevisiae
M. W. Briggs and J. S. Butler
Department of Microbiology and Immunology, University of Rochester School of Medicine and Dentistry, Rochester, New York 14624
We isolated spontaneous extragenic suppressors of a temperature-sensitive, lethal poly(A) polymerase mutation (pap1-1) in Saccharomyces cerevisiae that restore growth at the restrictive temperature of 30{deg}. Three of five suppressors represent alleles of the PDS2 complementation group. The recessive pds2-1 mutation exerts dominant allele-specific suppression over pap1-1, suggesting a direct functional interaction. The suppressor restores to near normal the steady-state concentrations of various mRNAs and total poly(A) reduced by pap1-1 at 30{deg}. Transcriptional chase experiments detect no reduction in the decay rates of mRNAs in the suppressor strain, suggesting that the restoration of steady-state message levels results from increased stable mRNA synthesis. Molecular cloning shows PDS2 to be allelic to RET1, which encodes the second-largest subunit of RNA polymerase III. We observe alterations in both the length and the steady-state amounts of RNA polymerase III transcripts in pds2-1 strains. Previously identified ret1 alleles do not suppress pap1-1, indicating that the pds2 alleles we isolated represent a specific class of RET1 mutations that suppress pap1-1. Suppression of pap1-1 by mutations in an RNA polymerase III subunit suggests a number of potentially novel interactions between these enzymes.
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