Editor: Roger Buxton
Mycobacterium tuberculosis Rv1302 and Mycobacterium smegmatis MSMEG___4947 have WecA function and MSMEG__4947 is required for the growth of M. smegmatis
Article first published online: 23 JUN 2010
© 2010 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved
FEMS Microbiology Letters
Volume 310, Issue 1, pages 54–61, September 2010
How to Cite
Jin, Y., Xin, Y., Zhang, W. and Ma, Y. (2010), Mycobacterium tuberculosis Rv1302 and Mycobacterium smegmatis MSMEG___4947 have WecA function and MSMEG__4947 is required for the growth of M. smegmatis. FEMS Microbiology Letters, 310: 54–61. doi: 10.1111/j.1574-6968.2010.02045.x
- Issue published online: 4 AUG 2010
- Article first published online: 23 JUN 2010
- Received 29 March 2010; revised 7 June 2010; accepted 8 June 2010.Final version published online 20 July 2010.
- GlcNAc-1-phosphate transferase;
The disaccharide d-N-acetylglucosamine-l-rhamnose plays an important role in the mycobacterial cell wall as a linker connecting arabinogalactan and peptidoglycan via a phosphodiester linkage. The first step of the disaccharide linker is the formation of decaprenyl phosphate-GlcNAc, which is catalyzed by GlcNAc-1-phosphate transferase. In Gram-negative bacteria, the wecA gene specifies the UDP-GlcNAc: undecaprenyl phosphate GlcNAc-1-phosphate transferase (WecA), which catalyzes the first step in the biosynthesis of lipopolysaccharide O-antigen. Mycobacterium tuberculosis Rv1302 and Mycobacterium smegmatis MSMEG_4947 show homology to Escherichia coli WecA protein. We cloned Rv1302 and MSMEG_4947 and introduced plasmids pYJ-1 (carrying Rv1302) and pYJ-2 (carrying MSMEG_4947) into a wecA-defective strain of E. coli MV501, respectively. Lipopolysaccharide analysis demonstrated that lipopolysaccharide synthesis in MV501 (pYJ-1) and MV501 (pYJ-2) was restored upon complementation with Rv1302 and MSMEG_4947, respectively. This provides the first evidence that Rv1302 and MSMEG_4947 have the same function as E. coli WecA. We also generated an M. smegmatis MSMEG_4947 knockout mutant using a homologous recombination strategy. The disruption of MSMEG_4947 in the M. smegmatis genome resulted in the loss of viability at a nonpermissive temperature. Scanning electron microscopy and transmission electron microscopy results showed that the lack of the MSMEG_4947 protein causes drastic morphological changes in M. smegmatis.