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PMID: 2649886 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Identification and purification of a single-stranded-DNA-specific exonuclease encoded by the recJ gene of Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 8 ·1989-04-00 ·Pages 2627-31

Lovett ST, Kolodner RD

Abstract

The Escherichia coli recJ gene product was overproduced using a plasmid that carries the recJ gene downstream of a strong regulatable promoter and a strong ribosome-binding site. Overexpression of recJ produced a concomitant increase in the levels of single-stranded-DNA-specific nuclease activity present in crude cell extracts. This nuclease activity was purified to homogeneity and found to reside in a 60-kDa polypeptide. This polypeptide was induced with recJ overexpression and had the size and N-terminal amino acid sequence identical to the predicted RecJ protein sequence. The RecJ nuclease degraded linear single-stranded DNA but did not have exonuclease activity on linear double-stranded substrates or endonuclease activity on either single-stranded or double-stranded substrates. The RecJ exonuclease had greater activity on duplex DNA molecules with 5'-rather than 3'-single-stranded tails.

MeSH Terms
Amino Acid Sequence Base Sequence DNA, Single-Stranded/metabolism Escherichia coli/enzymology,genetics Exodeoxyribonucleases/genetics,isolation & purification Gene Expression Regulation Genes, Bacterial Kinetics Molecular Sequence Data Mutation Recombination, Genetic Substrate Specificity
Chemicals
DNA, Single-Stranded Exodeoxyribonucleases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Lovett S T
Department of Cell and Molecular Biology, Dana-Farber Cancer Institute, Boston, MA 02115.
Kolodner R D
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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.86.8.2627
Published
1989-04-00
Pages
2627-31
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC286970
Subset
IM
Grants
NIGMS NIH HHS · GM26017 · United States
External Links
PubMed source DOI: 10.1073/pnas.86.8.2627 PMC Full Text View journal details
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