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PMID: 16522650 Published · epublish English Journal Article Research Support, Non-U.S. Gov't

Human replication protein A can suppress the intrinsic in vitro mutator phenotype of human DNA polymerase lambda.

Nucleic acids research ·Vol. 34 ·No. 5 ·2006-00-00 ·Pages 1405-15

Maga G, Shevelev I, Villani G, Spadari S, Hübscher U

Abstract

DNA polymerase lambda (pol lambda) is a member of the X family DNA polymerases and is endowed with multiple enzymatic activities. In this work we investigated the in vitro miscoding properties of full-length, human pol lambda either in the absence or in the presence of the human auxiliary proteins proliferating cell nuclear antigen (PCNA) and replication protein A (RP-A). Our data suggested that (i) pol lambda had an intrinsic ability to create mismatches and to incorporate ribonucleotides at nearly physiological Mn++ and Mg++ concentrations; (ii) the sequence of the template-primer could influence the misincorporation frequency of pol lambda; (iii) pol lambda preferentially generated G:T and G:G mismatches; (iv) RP-A, but not PCNA, selectively prevented misincorporation of an incorrect nucleotide by pol lambda, without affecting correct incorporation and (v) this inhibitory effect required a precise ratio between the concentrations of pol lambda and RP-A. Possible physiological implications of these findings for the in vivo fidelity of pol lambda are discussed.

MeSH Terms
Base Pair Mismatch DNA Polymerase beta/metabolism Deoxyribonucleotides/metabolism Humans Magnesium/chemistry Manganese/chemistry Phenotype Proliferating Cell Nuclear Antigen/physiology Replication Protein A/physiology Ribonucleotides/metabolism Templates, Genetic
Chemicals
Deoxyribonucleotides Proliferating Cell Nuclear Antigen RPA1 protein, human Replication Protein A Ribonucleotides Manganese DNA polymerase beta2 DNA Polymerase beta Magnesium
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Maga Giovanni
Istituto di Genetica Molecolare, IGM-CNR, Pavia, Italy. [email protected]
Shevelev Igor
Villani Giuseppe
Spadari Silvio
Hübscher Ulrich
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkl032
Email
[email protected]
Published
2006-00-00
Epub
2006-00-06
Pages
1405-15
Language
English
Region
England
NLM ID
0411011
PMCID
PMC1390690
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/gkl032 PMC Full Text View journal details
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