Characterization of an ATP-dependent DNA ligase encoded by Haemophilus influenzae

被引：47

作者：

Cheng, CH ^{[1
]}

Shuman, S ^{[1
]}

机构：

[1] SLOAN KETTERING INST, PROGRAM MOL BIOL, NEW YORK, NY 10021 USA

来源：

NUCLEIC ACIDS RESEARCH | 1997年 / 25卷 / 07期

关键词：

D O I：

10.1093/nar/25.7.1369

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 (K-m = 0.2 mu M) 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.

引用

页码：1369 / 1374

页数：6

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