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. 2004 Jul;186(14):4510-9.
doi: 10.1128/JB.186.14.4510-4519.2004.

Synthesis of the heteropolysaccharide O antigen of Escherichia coli O52 requires an ABC transporter: structural and genetic evidence

Lu Feng  1 , Sof'ya N Senchenkova, Jinghua Yang, Alexander S Shashkov, Jiang Tao, Hongjie Guo, Jiansong Cheng, Yi Ren, Yuriy A Knirel, Peter R Reeves, Lei Wang
Affiliations

Synthesis of the heteropolysaccharide O antigen of Escherichia coli O52 requires an ABC transporter: structural and genetic evidence

Lu Feng et al. J Bacteriol. 2004 Jul.

Abstract

The structural and genetic organization of the Escherichia coli O52 O antigen was studied. As identified by sugar and methylation analysis and nuclear magnetic resonance spectroscopy, the O antigen of E. coli O52 has a partially O-acetylated disaccharide repeating unit (O unit) containing D-fucofuranose and 6-deoxy-D-manno-heptopyranose, as well as a minor 6-deoxy-3-O-methylhexose (most likely, 3-O-methylfucose). The O-antigen gene cluster of E. coli O52, which is located between the galF and gnd genes, was found to contain putative genes for the synthesis of the O-antigen constituents, sugar transferase genes, and ABC-2 transporter genes. Further analysis confirmed that O52 employs an ATP-binding cassette (ABC) transporter-dependent pathway for translocation and polymerization of the O unit. This is the first report of an ABC transporter being involved in translocation of a heteropolysaccharide O antigen in E. coli. Genes specific for E. coli O52 were also identified.

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Figures

FIG. 1.
FIG. 1.
125-MHz 13C-NMR spectra of the O-deacetylated (A) and initial (B) O polysaccharide of E. coli O52.
FIG. 2.
FIG. 2.
O-antigen gene cluster of E. coli O52.
FIG. 3.
FIG. 3.
Deletion analysis of E. coli O52 orf16 (A) and wzm (B). Membrane extracts were electrophoresed on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels and stained by silver staining. (A) Lane 1, G1066 (E. coli O52 type strain); lane 2, H1044 (G1066 missing the orf16 gene); lane 3, G1066 (E. coli O52 type strain). (B) Lane 1, H1042 (G1066 missing the wzm gene); lane 2, H1043 (H1042 with plasmid pLW1001 carrying the E. coli O52 wzm gene); lane 3, G1066 (E. coli O52 type strain).
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