{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T04:47:40Z","timestamp":1790743660571,"version":"4.1.0"},"reference-count":56,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2017,4,15]],"date-time":"2017-04-15T00:00:00Z","timestamp":1492214400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Bacteriol"],"published-print":{"date-parts":[[2017,4,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            In Gram-negative bacteria, the peptidoglycan (PG) cell wall is a significant structural barrier for outer membrane protein assembly. In\n            <jats:named-content content-type=\"genus-species\">Aeromonas hydrophila<\/jats:named-content>\n            , outer membrane multimerization of the type II secretion system (T2SS) secretin ExeD requires the function of the inner membrane assembly factor complex ExeAB. The putative mechanism of the complex involves the reorganization of PG and localization of ExeD, whereby ExeA functions by interacting with PG to form a site for secretin assembly and ExeB forms an interaction with ExeD. This mechanism led us to hypothesize that increasing the pore size of PG would circumvent the requirement for ExeA in the assembly of the ExeD secretin. Growth of\n            <jats:named-content content-type=\"genus-species\">A. hydrophila<\/jats:named-content>\n            in 270 mM Gly reduced PG cross-links by approximately 30% and led to the suppression of secretin assembly defects in\n            <jats:italic>exeA<\/jats:italic>\n            strains and in those expressing ExeA mutants by enabling localization of the secretin in the outer membrane. We also established a heterologous ExeD assembly system in\n            <jats:named-content content-type=\"genus-species\">Escherichia coli<\/jats:named-content>\n            and showed that ExeAB and ExeC are the only\n            <jats:named-content content-type=\"genus-species\">A. hydrophila<\/jats:named-content>\n            proteins required for the assembly of the ExeD secretin in\n            <jats:named-content content-type=\"genus-species\">E. coli<\/jats:named-content>\n            and that ExeAB-independent assembly of ExeD can occur upon overexpression of the\n            <jats:sc>d<\/jats:sc>\n            ,\n            <jats:sc>d<\/jats:sc>\n            -carboxypeptidase PBP 5. These results support an assembly model in which, upon binding to PG, ExeA induces multimerization and pore formation in the sacculus, which enables ExeD monomers to interact with ExeB and assemble into a secretin that both is inserted in the outer membrane and crosses the PG layer to interact with the inner membrane platform of the T2SS.\n          <\/jats:p>\n          <jats:p>\n            <jats:bold>IMPORTANCE<\/jats:bold>\n            The PG layer imposes a strict structural impediment for the assembly of macromolecular structures that span the cell envelope and serve as virulence factors in Gram-negative species. This work revealed that by decreasing PG cross-linking by growth in Gly, the absolute requirement for the PG-binding activity of ExeA in the assembly of the ExeD secretin was alleviated in\n            <jats:named-content content-type=\"genus-species\">A. hydrophila<\/jats:named-content>\n            . In a heterologous assembly model in\n            <jats:named-content content-type=\"genus-species\">E. coli<\/jats:named-content>\n            , expression of the carboxypeptidase PBP 5 could relieve the requirement for ExeAB in the assembly of the ExeD secretin. These results provide some mechanistic details of the ExeAB assembly complex function, in which the PG-binding and oligomerization functions of ExeAB are used to create a pore in the PG that is required for secretin assembly.\n          <\/jats:p>","DOI":"10.1128\/jb.00617-16","type":"journal-article","created":{"date-parts":[[2017,1,31]],"date-time":"2017-01-31T02:50:34Z","timestamp":1485831034000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":12,"title":["Alterations in Peptidoglycan Cross-Linking Suppress the Secretin Assembly Defect Caused by Mutation of GspA in the Type II Secretion System"],"prefix":"10.1128","volume":"199","author":[{"given":"Elizabeth M.","family":"Vanderlinde","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, College of Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy G.","family":"Strozen","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, College of Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara B.","family":"Hern\u00e1ndez","sequence":"additional","affiliation":[{"name":"Laboratory for Molecular Infection Medicine Sweden, Department of Molecular Biology, Ume\u00e5 Center for Microbial Research, Ume\u00e5 University, Ume\u00e5, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Felipe","family":"Cava","sequence":"additional","affiliation":[{"name":"Laboratory for Molecular Infection Medicine Sweden, Department of Molecular Biology, Ume\u00e5 Center for Microbial Research, Ume\u00e5 University, Ume\u00e5, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S. 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