NAD+-dependent DNA ligases of Mycobacterium tuberculosis and Streptomyces coelicolor.
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.
Wilkinson
Adam
A
Phico Therapeutics, Ltd., Babraham Hall, Babraham, Cambridge, United Kingdom.
Sayer
Heather
H
Bullard
Desmond
D
Smith
Andrew
A
Day
Jonathan
J
Kieser
Tobias
T
Bowater
Richard
R
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D011088:DNA Ligases / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000201:enzymology / Q000235:genetics*; D009243:NAD / Q000378:metabolism*; D009841:Oligonucleotides / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics*; D013379:Substrate Specificity
D009841:Oligonucleotides; D009243:NAD; D011088:DNA Ligases