Identification and purification of a single-stranded-DNA-specific exonuclease encoded by the recJ gene of Escherichia coli - PubMed (original) (raw)
Identification and purification of a single-stranded-DNA-specific exonuclease encoded by the recJ gene of Escherichia coli
S T Lovett et al. Proc Natl Acad Sci U S A. 1989 Apr.
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.
Similar articles
- Orchestration of Haemophilus influenzae RecJ exonuclease by interaction with single-stranded DNA-binding protein.
Sharma R, Rao DN. Sharma R, et al. J Mol Biol. 2009 Feb 6;385(5):1375-96. doi: 10.1016/j.jmb.2008.11.041. Epub 2008 Nov 27. J Mol Biol. 2009. PMID: 19094995 - RecJ exonuclease: substrates, products and interaction with SSB.
Han ES, Cooper DL, Persky NS, Sutera VA Jr, Whitaker RD, Montello ML, Lovett ST. Han ES, et al. Nucleic Acids Res. 2006 Feb 18;34(4):1084-91. doi: 10.1093/nar/gkj503. Print 2006. Nucleic Acids Res. 2006. PMID: 16488881 Free PMC article. - RecQ helicase and RecJ nuclease provide complementary functions to resect DNA for homologous recombination.
Morimatsu K, Kowalczykowski SC. Morimatsu K, et al. Proc Natl Acad Sci U S A. 2014 Dec 2;111(48):E5133-42. doi: 10.1073/pnas.1420009111. Epub 2014 Nov 19. Proc Natl Acad Sci U S A. 2014. PMID: 25411316 Free PMC article. - Overexpression, purification and characterization of RecJ protein from Thermus thermophilus HB8 and its core domain.
Yamagata A, Masui R, Kakuta Y, Kuramitsu S, Fukuyama K. Yamagata A, et al. Nucleic Acids Res. 2001 Nov 15;29(22):4617-24. doi: 10.1093/nar/29.22.4617. Nucleic Acids Res. 2001. PMID: 11713311 Free PMC article. - Mutational analysis of the RecJ exonuclease of Escherichia coli: identification of phosphoesterase motifs.
Sutera VA Jr, Han ES, Rajman LA, Lovett ST. Sutera VA Jr, et al. J Bacteriol. 1999 Oct;181(19):6098-102. doi: 10.1128/JB.181.19.6098-6102.1999. J Bacteriol. 1999. PMID: 10498723 Free PMC article.
Cited by
- Prokaryotic DNA Crossroads: Holliday Junction Formation and Resolution.
Nautiyal A, Thakur M. Nautiyal A, et al. ACS Omega. 2024 Feb 27;9(11):12515-12538. doi: 10.1021/acsomega.3c09866. eCollection 2024 Mar 19. ACS Omega. 2024. PMID: 38524412 Free PMC article. Review. - RecJ3/4-aRNase J form a Ubl-associated nuclease complex functioning in survival against DNA damage in Haloferax volcanii.
Jia H, Dantuluri S, Margulies S, Smith V, Lever R, Allers T, Koh J, Chen S, Maupin-Furlow JA. Jia H, et al. mBio. 2023 Aug 31;14(4):e0085223. doi: 10.1128/mbio.00852-23. Epub 2023 Jul 17. mBio. 2023. PMID: 37458473 Free PMC article. - The Streptococcus agalactiae Exonuclease ExoVII Is Required for Resistance to Exogenous DNA-Damaging Agents.
Briaud P, Gautier T, Rong V, Mereghetti L, Lanotte P, Hiron A. Briaud P, et al. J Bacteriol. 2023 Jun 27;205(6):e0002423. doi: 10.1128/jb.00024-23. Epub 2023 May 10. J Bacteriol. 2023. PMID: 37162366 Free PMC article. - Mechanism of RecF-RecO-RecR cooperation in bacterial homologous recombination.
Nirwal S, Czarnocki-Cieciura M, Chaudhary A, Zajko W, Skowronek K, Chamera S, Figiel M, Nowotny M. Nirwal S, et al. Nat Struct Mol Biol. 2023 May;30(5):650-660. doi: 10.1038/s41594-023-00967-z. Epub 2023 Apr 20. Nat Struct Mol Biol. 2023. PMID: 37081315 - Beneficial and detrimental genes in the cellular response to replication arrest.
Schons-Fonseca L, Lazova MD, Smith JL, Anderson ME, Grossman AD. Schons-Fonseca L, et al. PLoS Genet. 2022 Dec 27;18(12):e1010564. doi: 10.1371/journal.pgen.1010564. eCollection 2022 Dec. PLoS Genet. 2022. PMID: 36574412 Free PMC article.
References
- Nature. 1970 Aug 15;227(5259):680-5 - PubMed
- Methods Enzymol. 1980;65(1):201-11 - PubMed
- Proc Natl Acad Sci U S A. 1971 Apr;68(4):824-7 - PubMed
- Methods Enzymol. 1983;101:20-78 - PubMed
- J Bacteriol. 1984 Jan;157(1):190-6 - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Molecular Biology Databases