Assembly of cyclic enterobacterial common antigen in Escherichia coli K-12.
نویسندگان
چکیده
We describe here the purification and quantification of a water-soluble cyclic form of enterobacterial common antigen (ECA(CYC)) 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 ECA(CYC) 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 microg ECA(CYC) per milligram (dry weight), and cell fractionation studies revealed that these molecules were localized exclusively in the periplasm. The synthesis and assembly of ECA(CYC) 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 ECA(CYC) was dependent on the wzzE gene, which is required for the modulation of the polysaccharide chain lengths of phosphoglyceride-linked ECA (ECA(PG)). The presence of ECA(CYC) 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 ECA(PG), ECA(CYC) may be synthesized in many, if not all, members of the Enterobacteriaceae.
منابع مشابه
The modality of enterobacterial common antigen polysaccharide chain lengths is regulated by o349 of the wec gene cluster of Escherichia coli K-12.
The assembly of the phosphoglyceride-linked form of enterobacterial common antigen (ECA(PG)) occurs by a mechanism that involves modulation of polysaccharide chain length. However, the genetic determinant of this modulation has not been identified. Site-directed mutagenesis of o349 of the Escherichia coli K-12 wec gene cluster revealed that this locus encodes a Wzz protein that specifically mod...
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متن کاملIdentification and biosynthesis of cyclic enterobacterial common antigen in Escherichia coli.
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ورودعنوان ژورنال:
- Journal of bacteriology
دوره 187 20 شماره
صفحات -
تاریخ انتشار 2005