PMID: 9421510Feb 28, 1998Paper

Chlorella virus DNA ligase: nick recognition and mutational analysis

Nucleic Acids Research
V Sriskanda, Stewart Shuman

Abstract

Chlorella virus PBCV-1 DNA ligase seals nicked DNA substrates consisting of a 5'-phosphate-terminated strand and a 3'-hydroxyl-terminated strand annealed to a bridging DNA template strand. The enzyme discriminates at the DNA binding step between substrates containing a 5'-phosphate versus a 5'-hydroxyl at the nick. Mutational analysis of the active site motif KxDGxR (residues 27-32) illuminates essential roles for the conserved Lys, Asp and Arg moieties at different steps of the ligase reaction. Mutant K27A is unable to form the covalent ligase-(Lys-straightepsilonN-P)-adenylate intermediate and hence cannot activate a nicked DNA substrate via formation of the DNA-adenylate intermediate. Nonetheless, K27A catalyzes phosphodiester bond formation at a pre-adenylated nick. This shows that the active site lysine is not required for the strand closure reaction. K27A binds to nicked DNA-adenylate, but not to a standard DNA nick. This suggests that occupancy of the AMP binding pocket of DNA ligase is important for nick recognition. Mutant D29A is active in enzyme-adenylate formation and binds readily to nicked DNA, but is inert in DNA-adenylate formation. R32A is unable to catalyze any of the three reactions of the ligation pathway an...Continue Reading

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Citations

Feb 13, 2001·Current Opinion in Biotechnology·J BanérU Landegren
Oct 20, 2005·Journal of Bioscience and Bioengineering·T YamadaS Hiramatsu
Jul 10, 2007·Nature Structural & Molecular Biology·Pravin A NairStewart Shuman
Dec 11, 2002·European Journal of Biochemistry·Alexei V Cherepanov, Simon de Vries
May 7, 2008·Proceedings of the National Academy of Sciences of the United States of America·Aurélien CrutNynke H Dekker
Nov 26, 2010·The Journal of Biological Chemistry·Anupam K Chakravarty, Stewart Shuman
Feb 22, 2011·The Journal of Biological Chemistry·Poulami Samai, Stewart Shuman
Apr 30, 2011·The Journal of Biological Chemistry·Poulami Samai, Stewart Shuman
Oct 27, 2011·The Journal of Biological Chemistry·Gregory J S LohmanThomas C Evans
Jan 22, 2008·The Journal of Biological Chemistry·Hui Zhu, Stewart Shuman
Jul 22, 1998·Nucleic Acids Research·V Sriskanda, S Shuman
Aug 24, 1999·Nucleic Acids Research·A J Doherty
Nov 1, 2000·Nucleic Acids Research·A J Doherty, S W Suh
Aug 26, 2004·Nucleic Acids Research·Pingfang LiuLawrence C Sowers
Oct 27, 2009·Nucleic Acids Research·Joohoon Kim, Milan Mrksich
Oct 30, 2009·Nucleic Acids Research·Kyu-Han KimKyu-Won Kim
Jul 20, 2010·Annual Review of Analytical Chemistry·Tim ConzeUlf Landegren
Jun 4, 2008·Annual Review of Biochemistry·Tom Ellenberger, Alan E Tomkinson
Aug 20, 2009·RNA·Naoko Tanaka, Stewart Shuman
Sep 22, 2011·PloS One·Maude Tremblay-LétourneauMartin Bisaillon
Nov 10, 2013·Nucleic Acids Research·Gregory J S LohmanThomas C Evans
Sep 14, 2002·Proceedings of the National Academy of Sciences of the United States of America·C Kiong Ho, Stewart Shuman
Feb 12, 2008·Current Opinion in Structural Biology·John M Pascal
Jun 1, 2006·DNA Repair·Caterina MarchettiPenny Jeggo
Oct 29, 2015·Archaea : an International Microbiological Journal·Maiko TanabeHirokazu Nishida
Jul 2, 2015·Nucleic Acids Research·Vandna KukshalTom Ellenberger
Oct 21, 2004·Molecular Cell·Jayakrishnan Nandakumar, Stewart Shuman
Nov 5, 1999·Annual Review of Microbiology·J L Van Etten, R H Meints
May 1, 2007·Molecular Cell·Jayakrishnan NandakumarStewart Shuman
Jul 18, 2006·Virology·Delphine Benarroch, Stewart Shuman
Feb 5, 2000·Journal of Molecular Biology·A J Doherty, T R Dafforn
Jan 8, 1999·Journal of Molecular Biology·D J Timson, D B Wigley
Jul 15, 2018·Nucleic Acids Research·Adele WilliamsonHanna-Kirsti S Leiros
Dec 26, 2001·The Journal of Biological Chemistry·Verl Sriskanda, Stewart Shuman

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