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A Yeast Gene Required for DNA Replication Encodes a Protein with Homology to DNA Helicases

Budd, Martin E. and Campbell, Judith L. (1995) 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, 92 (17). pp. 7642-7646. ISSN 0027-8424. PMCID PMC41201. doi:10.1073/pnas.92.17.7642.

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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.

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Additional Information:© 1995 by National Academy of Sciences Communicated by John Abelson, California Institute of Technology, Pasadena, CA, April 28, 1995 This work was supported by U.S. Public Health Service Grant GM25508. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
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Issue or Number:17
PubMed Central ID:PMC41201
Record Number:CaltechAUTHORS:BUDpnas95
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:3453
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Deposited On:08 Jun 2006
Last Modified:08 Nov 2021 19:56

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