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PMID: 10924459 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

POB3 is required for both transcription and replication in the yeast Saccharomyces cerevisiae.

Genetics ·Vol. 155 ·No. 4 ·2000-08-00 ·Pages 1593-606

Schlesinger MB, Formosa T

Abstract

Spt16 and Pob3 form stable heterodimers in Saccharomyces cerevisiae, and homologous proteins have also been purified as complexes from diverse eukaryotes. This conserved factor has been implicated in both transcription and replication and may affect both by altering the characteristics of chromatin. Here we describe the isolation and properties of a set of pob3 mutants and confirm that they have defects in both replication and transcription. Mutation of POB3 caused the Spt(-) phenotype, spt16 and pob3 alleles displayed severe synthetic defects, and elevated levels of Pob3 suppressed some spt16 phenotypes. These results are consistent with previous reports that Spt16 and Pob3 act in a complex that modulates transcription. Additional genetic interactions were observed between pob3 mutations and the genes encoding several DNA replication factors, including POL1, CTF4, DNA2, and CHL12. pob3 alleles caused sensitivity to the ribonucleotide reductase inhibitor hydroxyurea, indicating a defect in a process requiring rapid dNTP synthesis. Mutation of the S phase checkpoint gene MEC1 caused pob3 mutants to lose viability rapidly under restrictive conditions, revealing defects in a process monitored by Mec1. Direct examination of DNA contents by flow cytometry showed that S phase onset and progression were delayed when POB3 was mutated. We conclude that Pob3 is required for normal replication as well as for transcription.

MeSH Terms
Alleles Carrier Proteins/genetics,physiology Cell Cycle/genetics Cell Cycle Proteins/genetics,physiology Cell Nucleus/genetics Chromatin/metabolism DNA Replication/genetics Electrophoresis, Polyacrylamide Gel Flow Cytometry Hydroxyurea/pharmacology Mutagenesis Mutation Nucleic Acid Synthesis Inhibitors/pharmacology S Phase/genetics Saccharomyces cerevisiae/genetics,physiology Saccharomyces cerevisiae Proteins Temperature Time Factors Transcription Factors/metabolism Transcription, Genetic/genetics Transcriptional Elongation Factors
Chemicals
Carrier Proteins Cell Cycle Proteins Chromatin Nucleic Acid Synthesis Inhibitors POB3 protein, S cerevisiae SPT16 protein, S cerevisiae Saccharomyces cerevisiae Proteins Transcription Factors Transcriptional Elongation Factors Hydroxyurea
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Schlesinger M B
Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, Utah 84132, USA.
Formosa T
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Article Info
Journal
Genetics
Abbr.
Genetics
ISSN
0016-6731
DOI
10.1093/genetics/155.4.1593
Published
2000-08-00
Pages
1593-606
Language
English
Region
United States
NLM ID
0374636
PMCID
PMC1461200
Subset
IM
External Links
PubMed source DOI: 10.1093/genetics/155.4.1593 PMC Full Text View journal details
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