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Journal of Bacteriology, October 2005, p. 6917-6927, Vol. 187, No. 20
0021-9193/05/$08.00+0     doi:10.1128/JB.187.20.6917-6927.2005

Assembly of Cyclic Enterobacterial Common Antigen in Escherichia coli K-12

Junko Kajimura, Arifur Rahman, and Paul D. Rick*

Department of Biochemistry and Molecular Biology, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-4799

Received 6 May 2005/ Accepted 20 June 2005

We describe here the purification and quantification of a water-soluble cyclic form of enterobacterial common antigen (ECACYC) from Escherichia coli K-12 as well as information regarding its subcellular location and the genetic loci involved in its assembly. Structural characterization of purified ECACYC molecules obtained from E. coli K-12 revealed that they uniformly contained four trisaccharide repeat units, and they were substituted with from zero to four O-acetyl groups. Cells from overnight cultures contained approximately 2 µg ECACYC per milligram (dry weight), and cell fractionation studies revealed that these molecules were localized exclusively in the periplasm. The synthesis and assembly of ECACYC were found to require the wzxE and wzyE genes of the wec gene cluster. These genes encode proteins involved in the transmembrane translocation of undecaprenylpyrophosphate-linked ECA trisaccharide repeat units and the polymerization of trisaccharide repeat units, respectively. Surprisingly, synthesis of ECACYC was dependent on the wzzE gene, which is required for the modulation of the polysaccharide chain lengths of phosphoglyceride-linked ECA (ECAPG). The presence of ECACYC in extracts of several other gram-negative enteric organisms was also demonstrated; however, it was not detected in cell extracts of Pseudomonas aeruginosa. These data suggest that in addition to ECAPG, ECACYC may be synthesized in many, if not all, members of the Enterobacteriaceae.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814-4799. Phone: (301) 295-3418. Fax: (301) 295-3512. E-mail: rickp{at}usuhs.mil.

Journal of Bacteriology, October 2005, p. 6917-6927, Vol. 187, No. 20
0021-9193/05/$08.00+0     doi:10.1128/JB.187.20.6917-6927.2005




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