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PMID:23777429

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Citation

Hartmann, N and Büttner, D (2013) The inner membrane protein HrcV from Xanthomonas spp. is involved in substrate docking during type III secretion. Mol. Plant Microbe Interact. 26:1176-89

Abstract

Pathogenicity of the gram-negative plant-pathogenic bacterium Xanthomonas campestris pv. vesicatoria depends on a membrane-spanning type III secretion (T3S) system, which translocates effector proteins into eukaryotic host cells. In this study, we characterized the T3S system component HrcV, which is a member of the YscV/FlhA family of inner membrane proteins. HrcV consists of eight transmembrane helices and a cytoplasmic region (HrcVC). Mutant and protein-protein interaction studies showed that HrcVC is essential for protein function and binds to T3S substrates, including the early substrate HrpB2, the pilus protein HrpE, and effector proteins. Furthermore, HrcVC interacts with itself and with components and control proteins of the T3S apparatus. The interaction of HrcVC with HrpB2, HrpE, and T3S system components depends on amino acid residues in a conserved motif, designated flagella/hypersensitive response/invasion proteins export pore (FHIPEP), which is located in a cytoplasmic loop between transmembrane helix four and five of HrcV. Mutations in the FHIPEP motif abolish HrcV function but do not affect the interaction of HrcVC with effector proteins.

Links

PubMed Online version:10.1094/MPMI-01-13-0019-R

Keywords

Amino Acid Sequence; Bacterial Proteins/chemistry; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Bacterial Proteins/secretion; Bacterial Secretion Systems; Binding Sites; Capsicum/microbiology; Cell Membrane/metabolism; Conserved Sequence; Cytoplasm/metabolism; Models, Molecular; Molecular Sequence Data; Mutation; Plant Diseases/microbiology; Protein Interaction Domains and Motifs; Protein Interaction Mapping; Protein Transport; Sequence Alignment; Xanthomonas campestris/genetics; Xanthomonas campestris/metabolism; Xanthomonas campestris/pathogenicity

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

XANCP:Q8PB94

GO:0009306: protein secretion

ECO:0000314:

P

Figure 5 demonstrates the analysis of amino acid residues of the FHIPEP motif to protein–protein interactions. When HrcVR138A-c-Myc and HrcV D161A-c-Myc were incubated with GST or GST fusion proteins, both coeluted with GST-HpaC, GST-HrpE, and GST fusions of T3S effectors but were not detected in the eluates of GST-HrcV324-645, GST-HrcN, GST-HrcU255-357, GST- HpaB, and GST-HrpB2. The absence of detectable interactions of HrcVR138A-c-Myc and HrcVD161A-c-Myc with HpaB, HrpB2, and T3S system components explains the loss-of-function phenotypes and implies that these interactions are needed for protein function.

complete
CACAO 9978

See also

References

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