NAD+-dependent DNA ligases of Mycobacterium tuberculosis and Streptomyces coelicolor

Adam Wilkinson; Heather Sayer; Desmond Bullard; Andrew Smith; Jonathan Day; Tobias Kieser; Richard Bowater

doi:10.1002/prot.10361

NAD+-dependent DNA ligases of Mycobacterium tuberculosis and Streptomyces coelicolor

Adam Wilkinson
, Heather Sayer
, Desmond Bullard
, Andrew Smith
, Jonathan Day
, Tobias Kieser
, Richard Bowater

Research output: Contribution to journal › Article › peer-review

21 Citations (Scopus)

46 Downloads (Pure)

Abstract

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.

Original language	English
Pages (from-to)	321-326
Number of pages	6
Journal	Proteins: Structure Function and Bioinformatics
Volume	51
Issue number	3
DOIs	https://doi.org/10.1002/prot.10361
Publication status	Published - 21 Mar 2003

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SDG 3 Good Health and Well-being

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Cite this

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abstract = "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.",

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AU - Wilkinson, Adam

AU - Sayer, Heather

AU - Bullard, Desmond

AU - Smith, Andrew

AU - Day, Jonathan

AU - Kieser, Tobias

AU - Bowater, Richard

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AB - 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.

U2 - 10.1002/prot.10361

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