Factors necessary for the export process of colicin E1 across cytoplasmic membrane of Escherichia coli

Abstract

Factors necessary for the export process of colicin E1 across the cytoplasmic membrane of E. coli were investigated. .beta.-Galactosidase activities from gene fusions between the colicin E1 and lacZ genes were recovered in the inner membrane fraction of E. coli when the region containing the internal signal-like sequence of colicin E1 was present, but were found in the soluble fraction when the region was eliminated. The colicin E1 export was reduced upon insertion mutation in a gene that is located downstream from the colicin E1 gene in the same operon and responsible for mitomycin C-induced killing of the host cell. A frame shift mutation of the colicin E1 plasmid was constructed to direct the protein which lost the COOH-terminal 13 of original colicin E1 and was altered in 6 residues of the new COOH-terminal portion. The aberrant colicin E1 that was inducibly synthesized remained inside the cells. Colicin E1 is exported with the aid of a product of the downstream gene and that the COOH-terminal portion is necessary for the export. The binding of colicin E1 to the cytoplasmic membrane through the internal signal-like sequence may be a step in the protein export process.