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Characterization of an ATP-dependent DNA ligase encoded by Haemophilus influenzae.
Molecular Biology Program, Sloan-Kettering Institute, New York, NY 10021 USA.
We report that Haemophilus influenzae encodes a 268 amino acid ATP-dependent DNA ligase. The specificity of Haemophilus DNA ligase was investigated using recombinant protein produced in Escherichia coli. The enzyme catalyzed efficient strand joining on a singly nicked DNA in the presence of magnesium and ATP (Km = 0.2 microM). Other nucleoside triphosphates or deoxynucleoside triphosphates could not substitute for ATP. Haemophilus ligase reacted with ATP in the absence of DNA substrate to form a covalent ligase-adenylate intermediate. This nucleotidyl transferase reaction required a divalent cation and was specific for ATP. The Haemophilus enzyme is the first example of an ATP-dependent DNA ligase encoded by a eubacterial genome. It is also the smallest member of the covalent nucleotidyl transferase superfamily, which includes the bacteriophage and eukaryotic ATP-dependent polynucleotide ligases and the GTP-dependent RNA capping enzymes.
PMID: 9060431 [PubMed - indexed for MEDLINE]
PMCID: PMC146593
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Cited by 6 PubMed Central articles
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Characterization of Agrobacterium tumefaciens DNA ligases C and D.
Zhu H, Shuman S.
Nucleic Acids Res. 2007; 35(11):3631-45. Epub 2007 May 8.
[Nucleic Acids Res. 2007]
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Characterization of three new competence-regulated operons in Haemophilus influenzae.
VanWagoner TM, Whitby PW, Morton DJ, Seale TW, Stull TL.
J Bacteriol. 2004 Oct; 186(19):6409-21.
[J Bacteriol. 2004]
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Staphylococcus aureus DNA ligase: characterization of its kinetics of catalysis and development of a high-throughput screening compatible chemiluminescent hybridization protection assay.
Gul S, Brown R, May E, Mazzulla M, Smyth MG, Berry C, Morby A, Powell DJ.
Biochem J. 2004 Nov 1; 383(Pt. 3):551-9.
[Biochem J. 2004]
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