Journal article
Essential functions of the 32 kDa subunit of yeast replication protein A
Nucleic acids research, Vol.37(7), pp.2313-2326
04/2009
DOI: 10.1093/nar/gkp090
PMCID: PMC2673435
PMID: 19244309
Abstract
Replication protein A (RPA) is a heterotrimeric (70, 32 and 14 kDa subunits), single-stranded DNA-binding protein required for cellular DNA metabolism. All subunits of RPA are essential for life, but the specific functions of the 32 and 14 kDa subunits remains unknown. The 32 kDa subunit (RPA2) has multiple domains, but only the central DNA-binding domain (called DBD D) is essential for life in Saccharomyces cerevisiae. To define the essential function(s) of RPA2 in S. cerevisiae, a series of site-directed mutant forms of DBD D were generated. These mutant constructs were then characterized in vitro and in vivo. The mutations had minimal effects on the overall structure and activity of the RPA complex. However, several mutants were shown to disrupt crosslinking of RPA2 to DNA and to dramatically lower the DNA-binding affinity of a RPA2-containing subcomplex. When introduced into S. cerevisiae, all DBD D mutants were viable and supported normal growth rates and DNA replication. These findings indicate that RPA2-DNA interactions are not essential for viability and growth in S. cerevisiae. We conclude that DNA-binding activity of RPA2 is dispensable in yeast and that the essential function of DBD D is intra- and/or inter-protein interactions.
Details
- Title: Subtitle
- Essential functions of the 32 kDa subunit of yeast replication protein A
- Creators
- Anne M Dickson - 1Department of Biochemistry, University of Iowa College of Medicine, Iowa City, IA 52242-2600, USA and 2Novosibirsk Institute of Bioorganic Chemistry, Prospekt Lavrentiev 8, 630090 Novosibirsk, RussiaYulia Krasikova - 1Department of Biochemistry, University of Iowa College of Medicine, Iowa City, IA 52242-2600, USA and 2Novosibirsk Institute of Bioorganic Chemistry, Prospekt Lavrentiev 8, 630090 Novosibirsk, RussiaPavel Pestryakov - 1Department of Biochemistry, University of Iowa College of Medicine, Iowa City, IA 52242-2600, USA and 2Novosibirsk Institute of Bioorganic Chemistry, Prospekt Lavrentiev 8, 630090 Novosibirsk, RussiaOlga Lavrik - 1Department of Biochemistry, University of Iowa College of Medicine, Iowa City, IA 52242-2600, USA and 2Novosibirsk Institute of Bioorganic Chemistry, Prospekt Lavrentiev 8, 630090 Novosibirsk, RussiaMarc S Wold - 1Department of Biochemistry, University of Iowa College of Medicine, Iowa City, IA 52242-2600, USA and 2Novosibirsk Institute of Bioorganic Chemistry, Prospekt Lavrentiev 8, 630090 Novosibirsk, Russia
- Resource Type
- Journal article
- Publication Details
- Nucleic acids research, Vol.37(7), pp.2313-2326
- Publisher
- Oxford University Press
- DOI
- 10.1093/nar/gkp090
- PMID
- 19244309
- PMCID
- PMC2673435
- ISSN
- 0305-1048
- eISSN
- 1362-4962
- Language
- English
- Date published
- 04/2009
- Academic Unit
- Radiation Oncology; Biochemistry and Molecular Biology
- Record Identifier
- 9984024538402771
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