Exoenzyme S of Pseudomonas aeruginosa is secreted by a type III pathway

外酶 操纵子 生物 开放式参考框架 突变体 打开阅读框 铜绿假单胞菌 核酸序列 遗传学 转座子突变 基因 转座因子 分子生物学 微生物学 肽序列 细菌
作者
Timothy L. Yahr,Joanne Goranson,Dara W. Frank
出处
期刊:Molecular Microbiology [Wiley]
卷期号:22 (5): 991-1003 被引量:272
标识
DOI:10.1046/j.1365-2958.1996.01554.x
摘要

Exoenzyme S is an extracellular ADP‐ribosyltransferase of Pseudomonas aeruginosa . Transposon mutagenesis of P. aeruginosa 388 was used to identify genes required for exoenzyme S production. Five Tn 5 Tc insertion mutants were isolated which exhibited an exoenzyme S‐deficient phenotype (388::Tn 5 Tc 469, 550, 3453, 4885, and 5590). Mapping experiments demonstrated that 388::Tn 5 Tc 3453, 4885, and 5590 possessed insertions within a 5.0 kb Eco RI fragment that is not contiguous with the exoenzyme S trans ‐regulatory operon. 388::Tn 5 Tc 469 and 550 mapped to a region downstream of the trans ‐regulatory operon which has been previously shown to contain a promoter region that is co‐ordinately regulated with exoenzyme S synthesis. Nucleotide sequence analysis of a 7.2 kb region flanking the 388::Tn 5 Tc 469 and 550 insertions, identified 12 contiguous open reading frames (ORFs). Database searches indicated that the first ORF, ExsD, is unique. The other 11 ORFs demonstrated high homology to the YscB–L proteins of the yersiniaeYop type III export apparatus. RNase‐protection analysis of wild‐type and mutant strains indicated that exsD and pscB–L form an operon. To determine whether ExoS was exported by a type III mechanism, derivatives consisting of internal deletions or lacking amino‐ or carboxy‐terminal residues were expressed in P. aeruginosa . Deletion analyses indicated that the amino‐terminal nine residues are required for ExoS export. Combined data from mutagenesis, regulatory, expression, and sequence analyses provide strong evidence that P. aeruginosa possesses a type III secretion apparatus which is required for the export of exoenzyme S and potentially other co‐ordinately regulated proteins.
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