Novel prlA alleles defective in supporting staphylokinase processing in Escherichia coli.
A class of prlA (secY) alleles of Escherichia coli (prlA4-1 and prlA401) which specifically block the export of staphylokinase has been identified (T. Iino and T. Sako, J. Biol. Chem. 263:19077-19082, 1988; T. Sako and T. Iino, J. Bacteriol. 170:5389-5391, 1988). To determine more precisely the region in PrlA (SecY) effective for the blockage of processing of the staphylokinase precursor, additional prlA mutants which failed to support processing of the staphylokinase precursor were isolated. Two of the five mutant alleles isolated (secY121 and secY161) complemented the temperature sensitivity of a secY24 strain and had no detectable effect on the processing of endogenous secretory proteins of E. coli. In addition, a staphylokinase mutant having glycine in place of serine at position 17 in its signal sequence relieved the detrimental effect of these mutations. All of these characteristics indicate that these two alleles resemble the prlA4-1 and prlA401 alleles. On the other hand, the remaining three mutant alleles (secY47, secY105, and secY112) had no significant PrlA activity. The mutations of secY121 and secY161 were mapped very close to those of prlA4-1 and prlA401 in the presumed transmembrane segment 7 of PrlA. These results indicate that transmembrane segment 7 of PrlA plays a crucial role in the recognition of the staphylokinase signal sequence.
Sako
T
T
Yakult Central Institute for Microbiological Research, Tokyo, Japan.
D000483:Alleles; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism*; D001692:Biological Transport; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D005816:Genetic Complementation Test; D008666:Metalloendopeptidases / Q000235:genetics*; D009154:Mutation; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals / Q000378:metabolism*; D000069816:SEC Translocation Channels; D013329:Structure-Activity Relationship
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D021382:Protein Sorting Signals; D000069816:SEC Translocation Channels; C057841:SecY protein, E coli; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus