The gene coding for the Hrp pilus structural protein is required for type III secretion of Hrp and Avr proteins in Pseudomonas syringae pv. tomato.
Bacterial surface appendages called pili often are associated with DNA and/or protein transfer between cells. The exact function of pili in the transfer process is not understood and is a matter of considerable debate. The Hrp pilus is assembled by the Hrp type III protein secretion system of Pseudomonas syringae pv. tomato (Pst) strain DC3000. In this study, we show that the hrpA gene, which encodes the major subunit of the Hrp pilus, is required for secretion of putative virulence proteins, such as HrpW and AvrPto. In addition, the hrpA gene is required for full expression of genes that encode regulatory, secretion, and effector proteins of the type III secretion system. hrpA-mediated gene regulation apparently is through effect on the mRNA level of two previously characterized regulatory genes, hrpR and hrpS. Ectopic expression of the hrpRS gene operon restored gene expression, but not protein secretion, in the hrpA mutant. Three single amino acid mutations at the HrpA carboxyl terminus were identified that affect the secretion or regulatory function of the HrpA protein. These results define an essential role of the Hrp pilus structural gene in protein secretion and coordinate regulation of the type III secretion system in Pst DC3000.
Wei
W
W
Department of Energy Plant Research Laboratory, Genetics Graduate Program, Michigan State University, East Lansing, MI 48824, USA.
Plovanich-Jones
A
A
Deng
W L
WL
Jin
Q L
QL
Collmer
A
A
Huang
H C
HC
He
S Y
SY
D000596:Amino Acids; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D053487:DEAD-box RNA Helicases; D004268:DNA-Binding Proteins / Q000235:genetics*; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D016296:Mutagenesis; D009876:Operon; D018933:Pili, Sex; D011133:Polysaccharide-Lyases / Q000235:genetics* / Q000378:metabolism; D011549:Pseudomonas / Q000235:genetics*; D020365:RNA Helicases; D012329:RNA, Bacterial; D014157:Transcription Factors; D014158:Transcription, Genetic
D000596:Amino Acids; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C085856:HrpR protein, Pseudomonas syringae; D012329:RNA, Bacterial; D014157:Transcription Factors; C081695:avrPto protein, Pseudomonas syringae; C060789:hrpS protein, Pseudomonas syringae; C092662:hrpA protein, E coli; D053487:DEAD-box RNA Helicases; D020365:RNA Helicases; D011133:Polysaccharide-Lyases; C023911:pectate lyase