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NAD + ‐dependent DNA ligases of Mycobacterium tuberculosis and Streptomyces coelicolor
Author(s) -
Wilkinson Adam,
Sayer Heather,
Bullard Desmond,
Smith Andrew,
Day Jonathan,
Kieser Tobias,
Bowater Richard
Publication year - 2003
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/prot.10361
Subject(s) - streptomyces coelicolor , biology , nad+ kinase , dna ligase , mycobacterium tuberculosis , dna ligases , dna , microbiology and biotechnology , escherichia coli , gene , genetics , biochemistry , enzyme , mutant , tuberculosis , medicine , pathology
Sequencing of the genomes of Mycobacterium tuberculosis H37Rv and Streptomyces coelicolor A3(2) identified putative genes for an NAD + ‐dependent DNA ligase. We have cloned both open reading frames and overexpressed the protein products in Escherichia coli . In vitro biochemical assays confirm that each of these proteins encodes a functional DNA ligase that uses NAD + as its cofactor. Expression of either protein is able to complement E. coli GR501, which carries a temperature‐sensitive mutation in ligA . Thus, in vitro and in vivo analyses confirm predictions that ligA genes from M. tuberculosis and S. coelicolor are NAD + ‐dependent DNA ligases. Proteins 2003;51:321–326. © 2003 Wiley‐Liss, Inc.