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PMID: 7644470 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

A yeast gene required for DNA replication encodes a protein with homology to DNA helicases.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 92 ·No. 17 ·1995-08-15 ·Pages 7642-6

Budd ME, Campbell JL

Abstract

A yeast gene has been identified by screening for DNA replication mutants using a permeabilized cell replication assay. The mutant is temperature sensitive for growth and shows a cell cycle phenotype typical of DNA replication mutants. RNA synthesis is normal in the mutant but DNA synthesis ceases upon shift to the nonpermissive temperature. The DNA2 gene was cloned by complementation of the dna2ts gene phenotype. The gene is essential for viability. The gene encodes a 172-kDa protein with characteristic DNA helicase motifs. A hemagglutinin epitope-Dna2 fusion protein was prepared and purified by conventional and immunoaffinity chromatography. The purified protein is a DNA-dependent ATPase and has 3' to 5' DNA helicase activity specific for forked substrates. A nuclease activity that endonucleolytically cleaves DNA molecules having a single-stranded 5' tail adjacent to a duplex region copurifies through all steps with the fusion protein.

Related Genes
DNA2 TRP1 dna2<down>ts</down>
MeSH Terms
Adenosine Triphosphatases/biosynthesis,chemistry,genetics Amino Acid Sequence Base Sequence Cloning, Molecular DNA Helicases/biosynthesis,chemistry,genetics DNA Replication/genetics Genes, Fungal Genetic Complementation Test Molecular Sequence Data Oligodeoxyribonucleotides Phenotype Plasmids Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins Sequence Homology, Amino Acid
Chemicals
Oligodeoxyribonucleotides Saccharomyces cerevisiae Proteins Adenosine Triphosphatases DNA Helicases DNA2 protein, S cerevisiae
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Budd M E
Braun Laboratories, California Institute of Technology, Pasadena 91125, USA.
Campbell J L
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29 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.92.17.7642
Published
1995-08-15
Pages
7642-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC41201
Subset
IM
Grants
NIGMS NIH HHS · GM25508 · United States
Databases
GENBANK
L11229, U00027
External Links
PubMed source DOI: 10.1073/pnas.92.17.7642 PMC Full Text View journal details
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