The phosphatase activity of mammalian polynucleotide kinase takes precedence over its kinase activity in repair of single strand breaks.

Nucleic Acids Research
Caroline J Dobson, Sarah L Allinson

Abstract

The dual function mammalian DNA repair enzyme, polynucleotide kinase (PNK), facilitates strand break repair through catalysis of 5'-hydroxyl phosphorylation and 3'-phosphate dephosphorylation. We have examined the relative activities of the kinase and phosphatase functions of PNK using a novel assay, which allows the simultaneous characterization of both activities in processing nicks and gaps containing both 3'-phosphate and 5'-hydroxyl. Under multiple turnover conditions the phosphatase activity of the purified enzyme is significantly more active than its kinase activity. Consistent with this result, phosphorylation of the 5'-hydroxyl is rate limiting in cell extract mediated-repair of a nicked substrate. On characterizing the effects of individually mutating the two active sites of PNK we find that while site-directed mutagenesis of the kinase domain of PNK does not affect its phosphatase activity, disruption of the phosphatase domain also abrogates kinase function. This loss of kinase function requires the presence of a 3'-phosphate, but it need not be present in the same strand break as the 5'-hydroxyl. PNK preferentially binds 3'-phosphorylated substrates and DNA binding to the phosphatase domain blocks further DNA bindin...Continue Reading

References

Oct 15, 1975·European Journal of Biochemistry·H TeraokaK Tsukada
Oct 1, 1973·International Journal of Radiation Biology and Related Studies in Physics, Chemistry, and Medicine·T CoquerelleU Hagen
Dec 31, 1982·Biochemical and Biophysical Research Communications·B H Pheiffer, S B Zimmerman
Aug 22, 1983·FEBS Letters·Y Habraken, W G Verly
Oct 15, 1996·Proceedings of the National Academy of Sciences of the United States of America·S W YangH A Nash
Nov 5, 1997·The Journal of Biological Chemistry·A M ChagovetzB D Preston
Aug 14, 1999·The Journal of Biological Chemistry·F Karimi-BusheriM Weinfeld
Oct 12, 2000·Nucleic Acids Research·M D VodenicharovG G Poirier
May 4, 2001·The Journal of Biological Chemistry·L K Wang, S Shuman
Jul 12, 2002·The EMBO Journal·Li Kai WangStewart Shuman
Mar 22, 2003·The Journal of Biological Chemistry·Bryan D SmithRonald T Raines
Nov 26, 2003·Mutation Research·Grigory L DianovSarah L Allinson
Apr 22, 2004·Proceedings of the National Academy of Sciences of the United States of America·Aghdass Rasouli-NiaMichael Weinfeld
Jul 21, 2004·Molecular Cell·Lee WiederholdTapas K Hazra
Sep 24, 2004·The EMBO Journal·Christine Anne KochDaniel Durocher
Mar 8, 2005·Molecular Cell·Nina K BernsteinJ N Mark Glover

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Citations

Dec 16, 2011·Proceedings of the National Academy of Sciences of the United States of America·Nicolas CoquelleJ N Mark Glover
Dec 21, 2011·Proceedings of the National Academy of Sciences of the United States of America·Matthew J Schellenberg, R Scott Williams
Jan 3, 2012·Nucleic Acids Research·Nasser TahbazMichael Weinfeld
Mar 1, 2011·Trends in Biochemical Sciences·Michael WeinfeldJ N Mark Glover
Dec 8, 2009·DNA Repair·James R WhitesideSarah L Allinson
Aug 10, 2007·Chembiochem : a European Journal of Chemical Biology·Changbei MaXiaoyuan Lv
May 23, 2012·The FEBS Journal·Annegrit SeifriedAntje Gohla
Nov 10, 2015·Chemical Reviews·Yongxi ZhaoChunhai Fan
Jan 7, 2016·The Journal of Biological Chemistry·Vladimir ShafirovichNicholas E Geacintov
May 2, 2019·Neurology. Genetics·Ilona KalasovaKeith W Caldecott
Dec 18, 2020·PloS One·Luis Bermúdez-GuzmánAlejandro Leal

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Methods Mentioned

BETA
PCR
FCS
electrophoresis
glycosylases

Software Mentioned

ImageQuant
SigmaPlot

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