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RecJ exonuclease: substrates, products and interaction with SSB
The RecJ exonuclease from Escherichia coli degrades single-stranded DNA (ssDNA) in the 5′–3′ direction and participates in homologous recombination and mismatch repair. The experiments described here address RecJ's substrate requirements and reaction products. RecJ complexes on a variety of 5′...
Autores principales: | , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2006
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1373692/ https://www.ncbi.nlm.nih.gov/pubmed/16488881 http://dx.doi.org/10.1093/nar/gkj503 |
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author | Han, Eugene S. Cooper, Deani L. Persky, Nicole S. Sutera, Vincent A. Whitaker, Richard D. Montello, Melissa L. Lovett, Susan T. |
author_facet | Han, Eugene S. Cooper, Deani L. Persky, Nicole S. Sutera, Vincent A. Whitaker, Richard D. Montello, Melissa L. Lovett, Susan T. |
author_sort | Han, Eugene S. |
collection | PubMed |
description | The RecJ exonuclease from Escherichia coli degrades single-stranded DNA (ssDNA) in the 5′–3′ direction and participates in homologous recombination and mismatch repair. The experiments described here address RecJ's substrate requirements and reaction products. RecJ complexes on a variety of 5′ single-strand tailed substrates were analyzed by electrophoretic mobility shift in the absence of Mg(2+) ion required for substrate degradation. RecJ required single-stranded tails of 7 nt or greater for robust binding; addition of Mg(2+) confirmed that substrates with 5′ tails of 6 nt or less were poor substrates for RecJ exonuclease. RecJ is a processive exonuclease, degrading ∼1000 nt after a single binding event to single-strand DNA, and releases mononucleotide products. RecJ is capable of degrading a single-stranded tail up to a double-stranded junction, although products in such reactions were heterogeneous and RecJ showed a limited ability to penetrate the duplex region. RecJ exonuclease was equally potent on 5′ phosphorylated and unphosphorylated ends. Finally, DNA binding and nuclease activity of RecJ was specifically enhanced by the pre-addition of ssDNA-binding protein and we propose that this specific interaction may aid recruitment of RecJ. |
format | Text |
id | pubmed-1373692 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2006 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-13736922006-02-23 RecJ exonuclease: substrates, products and interaction with SSB Han, Eugene S. Cooper, Deani L. Persky, Nicole S. Sutera, Vincent A. Whitaker, Richard D. Montello, Melissa L. Lovett, Susan T. Nucleic Acids Res Article The RecJ exonuclease from Escherichia coli degrades single-stranded DNA (ssDNA) in the 5′–3′ direction and participates in homologous recombination and mismatch repair. The experiments described here address RecJ's substrate requirements and reaction products. RecJ complexes on a variety of 5′ single-strand tailed substrates were analyzed by electrophoretic mobility shift in the absence of Mg(2+) ion required for substrate degradation. RecJ required single-stranded tails of 7 nt or greater for robust binding; addition of Mg(2+) confirmed that substrates with 5′ tails of 6 nt or less were poor substrates for RecJ exonuclease. RecJ is a processive exonuclease, degrading ∼1000 nt after a single binding event to single-strand DNA, and releases mononucleotide products. RecJ is capable of degrading a single-stranded tail up to a double-stranded junction, although products in such reactions were heterogeneous and RecJ showed a limited ability to penetrate the duplex region. RecJ exonuclease was equally potent on 5′ phosphorylated and unphosphorylated ends. Finally, DNA binding and nuclease activity of RecJ was specifically enhanced by the pre-addition of ssDNA-binding protein and we propose that this specific interaction may aid recruitment of RecJ. Oxford University Press 2006 2006-02-18 /pmc/articles/PMC1373692/ /pubmed/16488881 http://dx.doi.org/10.1093/nar/gkj503 Text en © The Author 2006. Published by Oxford University Press. All rights reserved |
spellingShingle | Article Han, Eugene S. Cooper, Deani L. Persky, Nicole S. Sutera, Vincent A. Whitaker, Richard D. Montello, Melissa L. Lovett, Susan T. RecJ exonuclease: substrates, products and interaction with SSB |
title | RecJ exonuclease: substrates, products and interaction with SSB |
title_full | RecJ exonuclease: substrates, products and interaction with SSB |
title_fullStr | RecJ exonuclease: substrates, products and interaction with SSB |
title_full_unstemmed | RecJ exonuclease: substrates, products and interaction with SSB |
title_short | RecJ exonuclease: substrates, products and interaction with SSB |
title_sort | recj exonuclease: substrates, products and interaction with ssb |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1373692/ https://www.ncbi.nlm.nih.gov/pubmed/16488881 http://dx.doi.org/10.1093/nar/gkj503 |
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