NDLI: Transfer of the First Arabinofuranose Residue to Galactan Is Essential for Mycobacterium smegmatis Viability▿

Content Provider	PubMed Central
Author	Shi, Libin Zhou, Roukun Liu, Zhentong Lowary, Todd L. Seeberger, Peter H. Stocker, Bridget L. Crick, Dean C. Khoo, Kay-hooi Chatterjee, Delphi
Abstract	The mycobacterial arabinan is an elaborate component of the cell wall with multiple glycosyl linkages and no repeating units. In Mycobacterium spp., the Emb proteins (EmbA, EmbB, and EmbC) have been identified as putative mycobacterial arabinosyltransferases implicated in the biogenesis of the cell wall arabinan. Furthermore, it is now evident that the EmbA and EmbB proteins are involved in the assembly of the nonreducing terminal motif of arabinogalactan and EmbC is involved in transferring arabinose, perhaps in the early stage of arabinan synthesis in lipoarabinomannan. It has also been shown that the Emb proteins are a target of the antimycobacterial drug ethambutol (EMB). In the search for additional mycobacterial arabinosyltransferases in addition to the Emb proteins, we disrupted MSMEG_6386 (an orthologue of Rv3792 and a gene upstream of embC) in Mycobacterium smegmatis. Allelic exchange at the chromosomal MSMEG_6386 locus of M. smegmatis could only be achieved in the presence of a rescue plasmid carrying a functional copy of MSMEG_6386 or Rv3792, strongly suggesting that MSMEG_6386 is essential. An in vitro arabinosyltransferase assay using a membrane preparation from M. smegmatis expressing Rv3792 and synthetic β-d-Galf-(1→5)-β-d-Galf-(1→6)-β-d-Galf-octyl and β-d-Galf-(1→6)-β-d-Galf-(1→5)-β-d-Galf-octyl showed that Rv3792 gene product can transfer an arabinose residue to the C-5 position of the internal 6-linked galactose. The reactions were insensitive to EMB, and when α-d-Manp-(1→6)-α-d-Manp-(1→6)-α-d-Manp-octylthiomethyl was used as an acceptor, no product was formed. These observations indicate that transfer of the first arabinofuranose residue to galactan is essential for M. smegmatis viability.
Related Links	http://dx.doi.org/10.1128/jb.00028-08
Ending Page	5255
Page Count	8
Starting Page	5248
File Format	PDF
ISSN	00219193
e-ISSN	10985530
Journal	Journal of Bacteriology
Issue Number	15
Volume Number	190
Language	English
Publisher	American Society for Microbiology (ASM)
Publisher Date	2008-08-01
Access Restriction	Open
Rights Holder	American Society for Microbiology (ASM)
Subject Keyword	Molecular Biology Microbiology Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Microbiology

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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