Intragenic and Extragenic Suppressors of Mutations in the Heptapeptide Repeat Domain of Saccharomyces cerevisiae RNA Polymerase II

INVESTIGATIONS

Intragenic and Extragenic Suppressors of Mutations in the Heptapeptide Repeat Domain of Saccharomyces cerevisiae RNA Polymerase II

M. L. Nonet and R. A. Young
Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, and Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

The largest subunit of RNA polymerase II contains a repeated heptapeptide sequence at its carboxy terminus. Yeast mutants with certain partial deletions of the carboxy-terminal repeat (CTR) domain are temperature-sensitive, cold-sensitive and are inositol auxotrophs. Intragenic and extragenic suppressors of the cold-sensitive phenotype of CTR domain deletion mutants were isolated and studied to investigate the function of this domain. Two types of intragenic suppressing mutations suppress the temperature-sensitivity, cold-sensitivity and inositol auxotrophy of CTR domain deletion mutants. Most intragenic mutations enlarge the repeat domain by duplicating various portions of the repeat coding sequence. Other intragenic suppressing mutations are point mutations in a conserved segment of the large subunit. An extragenic suppressing mutation (SRB2-1) was isolated that strongly suppresses the conditional and auxotrophic phenotypes of CTR domain mutations. The SRB2 gene was isolated and mapped, and an SRB2 partial deletion mutation (srb2{Delta}10) was constructed. The srb2{Delta}10 mutants are temperature-sensitive, cold-sensitive and are inositol auxotrophs. These phenotypes are characteristic of mutations in genes encoding components of the transcription apparatus. We propose that the SRB2 gene encodes a factor that is involved in RNA synthesis and may interact with the CTR domain of the large subunit of RNA polymerase II.

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				J. S. Kang, S. H. Kim, M. S. Hwang, S. J. Han, Y. C. Lee, and Y.-J. Kim The Structural and Functional Organization of the Yeast Mediator Complex J. Biol. Chem., November 2, 2001; 276(45): 42003 - 42010. [Abstract] [Full Text] [PDF]

				D. L. Lindstrom and G. A. Hartzog Genetic Interactions of Spt4-Spt5 and TFIIS With the RNA Polymerase II CTD and CTD Modifying Enzymes in Saccharomyces cerevisiae Genetics, October 1, 2001; 159(2): 487 - 497. [Abstract] [Full Text] [PDF]

				M. K. Shirra, J. Patton-Vogt, A. Ulrich, O. Liuta-Tehlivets, S. D. Kohlwein, S. A. Henry, and K. M. Arndt Inhibition of Acetyl Coenzyme A Carboxylase Activity Restores Expression of the INO1 Gene in a snf1 Mutant Strain of Saccharomyces cerevisiae Mol. Cell. Biol., September 1, 2001; 21(17): 5710 - 5722. [Abstract] [Full Text] [PDF]

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