The Cpx proteins of Escherichia coli K12. Immunologic detection of the chromosomal cpxA gene product.
Previous studies described lacZ'- and cat'-'cpxA fusion genes whose expression restored to normal all the phenotypic defects associated with cpxA mutations (Albin, R., and Silverman, P. M. (1984) Mol. Gen. Genet. 197, 272-279). Here, we show by DNA nucleotide sequence analysis that the fusion genes encode 241 carboxyl-terminal amino acids of the CpxA polypeptide. Using this information, we constructed a fusion gene containing the same 241 cpxA codons preceded by 1007 codons of beta-galactosidase. The resultant hybrid polypeptide was purified and used to raise an anti-(CpxA polypeptide) antiserum. Using the antiserum, we have identified the chromsomal Escherichia coli K12 cpxA gene product as a 52-kDa polypeptide. The polypeptide showed temperature-sensitive accumulation in a strain carrying both the cpxA2[Ts] and cpxB1 alleles and accumulated to a level higher than normal in cells that carried a high-copy number, cpxA+ plasmid. Immune precipitates of in vitro transcription-translation reactions with cpxA+ plasmids as template also contained a 52-kDa polypeptide, indistinguishable in electrophoretic mobility from the immunoreactive polypeptide synthesized in vivo. Two regions of amino acid sequence at the carboxyl-terminus of the CpxA polypeptide are significantly homologous to corresponding regions of the E. coli K12 EnvZ polypeptide, an inner membrane component that, like the CpxA polypeptide, is required to maintain the protein composition of the cell envelope. The cpxA coding sequence is followed by two repetitive extragenic palindrome sequences in opposite orientation.
D000595:Amino Acid Sequence; D000936:Antigen-Antibody Complex / Q000032:analysis; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics*; D001483:Base Sequence; D002876:Chromosomes, Bacterial / Q000502:physiology; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D005796:Genes; D005798:Genes, Bacterial; D007106:Immune Sera; D010957:Plasmids; D013696:Temperature
D000936:Antigen-Antibody Complex; D001426:Bacterial Proteins; D007106:Immune Sera; D004262:DNA Restriction Enzymes