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Synthesis of the Heteropolysaccharide O Antigen of Escherichia coli O52 Requires an ABC Transporter: Structural and Genetic Evidence

Lu Feng,^1,2, Sof'ya N. Senchenkova,^3, Jinghua Yang,^1, Alexander S. Shashkov,³ Jiang Tao,¹ Hongjie Guo,^1,2 Jiansong Cheng,¹ Yi Ren,¹ Yuriy A. Knirel,³ Peter R. Reeves,⁴ and Lei Wang^1,2,5*

TEDA School of Biological Sciences and Biotechnology,¹ Tianjin State Laboratory of Microbial Functional Genomics, TEDA College, Nankai University,² Tianjin Biochip Technology Corporation, TEDA, Tianjin 300457, People's Republic of China,⁵ N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow 119991, Russian Federation,³ School of Molecular and Microbial Biosciences (G08), University of Sydney, Sydney, North South Wales 2006, Australia⁴

Received 16 January 2004/ Accepted 12 April 2004

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.

L.F, S.N.S., and J.Y. contributed equally to this work.

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