| Characterization of an ATP-regulated DNA-processing enzyme and thermotolerant phosphoesterase in the radioresistant bacterium Deinococcus radiodurans. | |
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MedLine Citation:
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PMID: 20658964 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A multiprotein DNA-processing complex identified from Deinococcus radiodurans exhibits uncharacterized ATP-sensitive nuclease functions. DR0505 was one of the 24 polypeptides identified from the complex. It contains two 5' nucleotidase motifs, one is at the C-terminal end of the N-terminal CPDD (calcineurin phosphodiesterase domain), with the second at the C-terminal end of the protein. Recombinant DR0505 showed both phosphomonoesterase and phosphodiesterase activities with chromogenic substrates, showing higher affinity for bis-(p-nitrophenyl) phosphate than for p-nitrophenyl phosphate. The enzyme exhibited pH optima ranging from 8.0 to 9.0 and metal-ion-dependent thermotolerance of esterase functions. Both mono- and di-esterase activities were stable at temperatures up to 50 °C in the presence of Mg2+, whereas monoesterase activity was observed at temperatures up to 80 °C in the presence of Mn2+ and up to 50 °C with Ca2+. The purified enzyme showed 5' nucleotidase activity on a wide range of natural mononucleotides including cyclic mononucleotides and 8-oxo-GMP. DR0505 showed a nearly 7-fold higher activity on ADP than AMP, but this activity was inhibited with ATP. Interestingly, DR0505 also showed single-stranded endonuclease and 3'→5' exonuclease activities on both single-stranded and double-stranded DNA-substrates. Unlike for the exonuclease activity, the single-stranded endonuclease activities observed on stem-loop substrates and at the single strand-double-strand junction in forked-hairpin substrates were not inhibited with ATP. These results suggested that DR0505 is an ATP-regulated DNA-processing enzyme and a thermotolerant esterase in vitro. We therefore suggest possible roles of this enzyme in nucleotide recycling and DNA processing, which is required for efficient double-strand break repair and the high radiation tolerance observed in D. radiodurans. |
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Authors:
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Swathi Kota; C Vijaya Kumar; Hari S Misra |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: The Biochemical journal Volume: 431 ISSN: 1470-8728 ISO Abbreviation: Biochem. J. Publication Date: 2010 Oct |
Date Detail:
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Created Date: 2010-09-14 Completed Date: 2010-11-03 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 2984726R Medline TA: Biochem J Country: England |
Other Details:
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Languages: eng Pagination: 149-57 Citation Subset: IM |
Affiliation:
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Molecular Biology Division, Bhabha Atomic Research Centre, Mumbai 400 085, India. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Adenosine Triphosphate
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metabolism Bacterial Proteins / chemistry, metabolism* DNA Repair DNA Repair Enzymes / chemistry, metabolism* DNA, Bacterial / chemistry, metabolism* DNA-Binding Proteins / chemistry, metabolism Deinococcus / enzymology*, metabolism Gamma Rays Nucleotidases / chemistry, metabolism* Radiation Tolerance |
| Chemical | |
Reg. No./Substance:
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0/Bacterial Proteins; 0/DNA, Bacterial; 0/DNA-Binding Proteins; 56-65-5/Adenosine Triphosphate; EC 3.1.3.-/Nucleotidases; EC 6.5.1.-/DNA Repair Enzymes |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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