PMID: 2495026Apr 12, 1989Paper

Partial purification and characterization of an endonuclease from spinach that cleaves ultraviolet light-damaged duplex DNA

Biochimica Et Biophysica Acta
P W DoetschM R Valenzuela

Abstract

An endonuclease that cleaves ultraviolet light (UV)-damaged, supercoiled plasmid DNA was partially purified from spinach leaves (Spinacia oleracea) by a series of column chromatography steps. Dialysis of the enzyme against EDTA resulted in a greater than 90% loss of activity which could be fully restored following the addition of Zn2+, suggesting that divalent cations are associated with the active enzyme. The spinach endonuclease cleaved duplex, UV-damaged, end-labelled DNA of defined sequence at positions of adenine in the presence of salt (KH2PO4 or NaCl) concentrations of 50 mM or higher. Cleavage of UV-irradiated DNA was dose-dependent and increased steadily within a fluence range of 50-10,000 J/m2. The UV damage requirement and adenine cleavage specificity could be eliminated with lower salt concentrations (0-25 mM), suggesting that the endonuclease recognizes and incises single-stranded DNA. The properties of this enzyme, which we have termed nuclease SP, suggest that it may mediate a role in DNA repair and/or recombination processes in spinach.

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Citations

Aug 1, 1998·Mutation Research·E J VonarxB A Kunz
Dec 18, 2001·FEMS Microbiology Reviews·E S Rangarajan, V Shankar
May 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·J LenzP W Doetsch
Jul 1, 1995·Plant Physiology·A B Britt
Jun 1, 1996·Annual Review of Plant Physiology and Plant Molecular Biology·Anne B. Britt
Feb 15, 2003·FEMS Microbiology Reviews·Neelam A Desai, Vepatu Shankar
Jan 1, 1995·Critical Reviews in Microbiology·S U Gite, V Shankar
Oct 18, 1991·Biochemistry·J A StricklandP W Doetsch
Feb 9, 1993·Biochemistry·M CzuprynB L Vallee
Jun 29, 2011·Critical Reviews in Microbiology·Amruta Pramod Joshi, Sumedha Sharad Deshmukh

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