Structure-Function Analysis of the HrpB2-HrcU Interaction in the Xanthomonas citri Type III Secretion System
Identifieur interne : 001672 ( Main/Merge ); précédent : 001671; suivant : 001673Structure-Function Analysis of the HrpB2-HrcU Interaction in the Xanthomonas citri Type III Secretion System
Auteurs : Paola A. Cappelletti [Brésil] ; Rafael Freitas Dos Santos [Brésil] ; Alexandre M. Do Amaral [Brésil] ; Rafael Augusto Homem [Brésil] ; Thaís Dos Santos Souza [Brésil] ; Marcos A. Machado [Brésil] ; Chuck S. Farah [Brésil]Source :
- PLoS ONE [ 1932-6203 ] ; 2011.
Abstract
Bacterial type III secretion systems deliver protein virulence factors to host cells. Here we characterize the interaction between HrpB2, a small protein secreted by the
Url:
DOI: 10.1371/journal.pone.0017614
PubMed: 21408079
PubMed Central: 3052322
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Cappelletti</name><affiliation wicri:level="4"><nlm:aff id="aff1"><addr-line>Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, São Paulo, Brazil</addr-line></nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, São Paulo</wicri:regionArea><placeName><settlement type="city">São Paulo</settlement><region type="state">État de São Paulo</region></placeName><orgName type="university">Université de São Paulo</orgName></affiliation></author><author><name sortKey="Dos Santos, Rafael Freitas" sort="Dos Santos, Rafael Freitas" uniqKey="Dos Santos R" first="Rafael Freitas" last="Dos Santos">Rafael Freitas Dos Santos</name><affiliation wicri:level="4"><nlm:aff id="aff1"><addr-line>Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, São Paulo, Brazil</addr-line></nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, São Paulo</wicri:regionArea><placeName><settlement type="city">São Paulo</settlement><region type="state">État de São Paulo</region></placeName><orgName type="university">Université de São Paulo</orgName></affiliation></author><author><name sortKey="Do Amaral, Alexandre M" sort="Do Amaral, Alexandre M" uniqKey="Do Amaral A" first="Alexandre M." last="Do Amaral">Alexandre M. Do Amaral</name><affiliation wicri:level="1"><nlm:aff id="aff2"><addr-line>EMBRAPA Recursos Genéticos e Biotecnologia, Brasília, Distrito Federal, Brazil</addr-line></nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>EMBRAPA Recursos Genéticos e Biotecnologia, Brasília, Distrito Federal</wicri:regionArea><wicri:noRegion>Distrito Federal</wicri:noRegion></affiliation><affiliation wicri:level="3"><nlm:aff id="aff3"><addr-line>Centro APTA Citros “Sylvio Moreira”/IAC, Cordeiropolis, São Paulo, Brazil</addr-line></nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Centro APTA Citros “Sylvio Moreira”/IAC, Cordeiropolis, São Paulo</wicri:regionArea><placeName><settlement type="city">São Paulo</settlement><region type="state">État de São Paulo</region></placeName></affiliation></author><author><name sortKey="Homem, Rafael Augusto" sort="Homem, Rafael Augusto" uniqKey="Homem R" first="Rafael Augusto" last="Homem">Rafael Augusto Homem</name><affiliation wicri:level="3"><nlm:aff id="aff3"><addr-line>Centro APTA Citros “Sylvio Moreira”/IAC, Cordeiropolis, São Paulo, Brazil</addr-line></nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Centro APTA Citros “Sylvio Moreira”/IAC, Cordeiropolis, São Paulo</wicri:regionArea><placeName><settlement type="city">São Paulo</settlement><region type="state">État de São Paulo</region></placeName></affiliation></author><author><name sortKey="Dos Santos Souza, Thais" sort="Dos Santos Souza, Thais" uniqKey="Dos Santos Souza T" first="Thaís" last="Dos Santos Souza">Thaís Dos Santos Souza</name><affiliation wicri:level="4"><nlm:aff id="aff1"><addr-line>Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, São Paulo, Brazil</addr-line></nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, São Paulo</wicri:regionArea><placeName><settlement type="city">São Paulo</settlement><region type="state">État de São Paulo</region></placeName><orgName type="university">Université de São Paulo</orgName></affiliation></author><author><name sortKey="Machado, Marcos A" sort="Machado, Marcos A" uniqKey="Machado M" first="Marcos A." last="Machado">Marcos A. Machado</name><affiliation wicri:level="3"><nlm:aff id="aff3"><addr-line>Centro APTA Citros “Sylvio Moreira”/IAC, Cordeiropolis, São Paulo, Brazil</addr-line></nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Centro APTA Citros “Sylvio Moreira”/IAC, Cordeiropolis, São Paulo</wicri:regionArea><placeName><settlement type="city">São Paulo</settlement><region type="state">État de São Paulo</region></placeName></affiliation></author><author><name sortKey="Farah, Chuck S" sort="Farah, Chuck S" uniqKey="Farah C" first="Chuck S." last="Farah">Chuck S. Farah</name><affiliation wicri:level="4"><nlm:aff id="aff1"><addr-line>Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, São Paulo, Brazil</addr-line></nlm:aff><country xml:lang="fr">Brésil</country><wicri:regionArea>Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, São Paulo</wicri:regionArea><placeName><settlement type="city">São Paulo</settlement><region type="state">État de São Paulo</region></placeName><orgName type="university">Université de São Paulo</orgName></affiliation></author></analytic><series><title level="j">PLoS ONE</title><idno type="eISSN">1932-6203</idno><imprint><date when="2011">2011</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Bacterial type III secretion systems deliver protein virulence factors to host cells. Here we characterize the interaction between HrpB2, a small protein secreted by the <italic>Xanthomonas citri</italic> subsp. citri type III secretion system, and the cytosolic domain of the inner membrane protein HrcU, a paralog of the flagellar protein FlhB. We show that a recombinant fragment corresponding to the C-terminal cytosolic domain of HrcU produced in <italic>E. coli</italic> suffers cleavage within a conserved Asn264-Pro265-Thr266-His267 (NPTH) sequence. A recombinant HrcU cytosolic domain with N264A, P265A, T266A mutations at the cleavage site (HrcU<sub>AAAH</sub>) was not cleaved and interacted with HrpB2. Furthermore, a polypeptide corresponding to the sequence following the NPTH cleavage site also interacted with HrpB2 indicating that the site for interaction is located after the NPTH site. Non-polar deletion mutants of the <italic>hrcU</italic> and <italic>hrpB2</italic> genes resulted in a total loss of pathogenicity in susceptible citrus plants and disease symptoms could be recovered by expression of HrpB2 and HrcU from extrachromossomal plasmids. Complementation of the <italic>ΔhrcU</italic> mutant with HrcU<sub>AAAH</sub> produced canker lesions similar to those observed when complemented with wild-type HrcU. HrpB2 secretion however, was significantly reduced in the <italic>ΔhrcU</italic> mutant complemented with HrcU<sub>AAAH,</sub> suggesting that an intact and cleavable NPTH site in HrcU is necessary for total functionally of T3SS in <italic>X. citri</italic> subsp. citri. Complementation of the <italic>ΔhrpB2 X. citri</italic> subsp. citri strain with a series of <italic>hrpB2</italic> gene mutants revealed that the highly conserved HrpB2 C-terminus is essential for T3SS-dependent development of citrus canker symptoms <italic>in planta</italic>.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Blocker, A" uniqKey="Blocker A">A Blocker</name></author><author><name sortKey="Komoriya, K" uniqKey="Komoriya K">K Komoriya</name></author><author><name sortKey="Aizawa, S" uniqKey="Aizawa S">S Aizawa</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="He, Sy" uniqKey="He S">SY He</name></author><author><name sortKey="Nomura, K" uniqKey="Nomura K">K Nomura</name></author><author><name sortKey="Whittam, Ts" uniqKey="Whittam T">TS Whittam</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Buttner, D" uniqKey="Buttner D">D 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Mistry</name></author><author><name sortKey="Tate, J" uniqKey="Tate J">J Tate</name></author><author><name sortKey="Coggill, P" uniqKey="Coggill P">P Coggill</name></author><author><name sortKey="Heger, A" uniqKey="Heger A">A Heger</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Crooks, Ge" uniqKey="Crooks G">GE Crooks</name></author><author><name sortKey="Hon, G" uniqKey="Hon G">G Hon</name></author><author><name sortKey="Chandonia, Jm" uniqKey="Chandonia J">JM Chandonia</name></author><author><name sortKey="Brenner, Se" uniqKey="Brenner S">SE Brenner</name></author></analytic></biblStruct></listBibl></div1></back></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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