{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T18:53:00Z","timestamp":1768503180115,"version":"3.49.0"},"posted":{"date-parts":[[2023,1,10]]},"group-title":"Microbiology","reference-count":31,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,1,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  The GspD secretin is the outer membrane channel of the bacterial type II secretion system (T2SS) which secrets diverse effector proteins or toxins that cause severe diseases such as diarrhea and cholera. GspD needs to translocate from the inner to the outer membrane to exert its function, and this process is an essential step for T2SS to assemble. Here, we investigate two types of secretins discovered so far in\n                  <jats:italic>Escherichia coli<\/jats:italic>\n                  , GspD\n                  <jats:sub>\u03b1<\/jats:sub>\n                  and GspD\n                  <jats:sub>\u03b2<\/jats:sub>\n                  , respectively. By electron cryotomography subtomogram averaging, we determine\n                  <jats:italic>in situ<\/jats:italic>\n                  structures of all the key intermediate states of GspD\n                  <jats:sub>\u03b1<\/jats:sub>\n                  and GspD\n                  <jats:sub>\u03b2<\/jats:sub>\n                  in the translocation process, with resolution ranging from 9 \u00c5 to 19 \u00c5. In our results, GspD\n                  <jats:sub>\u03b1<\/jats:sub>\n                  and GspD\n                  <jats:sub>\u03b2<\/jats:sub>\n                  present entirely different membrane interaction patterns and ways of going across the peptidoglycan layer. We propose two distinct models for the membrane translocation of GspD\n                  <jats:sub>\u03b1<\/jats:sub>\n                  and GspD\n                  <jats:sub>\u03b2<\/jats:sub>\n                  , providing a comprehensive perspective on the inner to outer membrane biogenesis of T2SS secretins.\n                <\/jats:p>","DOI":"10.1101\/2023.01.10.523476","type":"posted-content","created":{"date-parts":[[2023,1,10]],"date-time":"2023-01-10T23:05:10Z","timestamp":1673391910000},"source":"Crossref","is-referenced-by-count":0,"title":["<i>In situ<\/i>\n                  structures of secretins from bacterial type II secretion system reveal their membrane interactions and translocation process"],"prefix":"10.64898","author":[{"given":"Zhili","family":"Yu","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yaoming","family":"Wu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Muyuan","family":"Chen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tong","family":"Huo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wei","family":"Zheng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Steven J.","family":"Ludtke","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaodong","family":"Shi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4897-9986","authenticated-orcid":false,"given":"Zhao","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2023011217451315000_2023.01.10.523476v1.1","doi-asserted-by":"crossref","unstructured":"Costa, T. R. D. et al. Secretion systems in Gram-negative bacteria: structural and mechanistic insights. Nat Rev Microbiol 13, rmicro3456 (2015).","DOI":"10.1038\/nrmicro3456"},{"key":"2023011217451315000_2023.01.10.523476v1.2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro2762"},{"key":"2023011217451315000_2023.01.10.523476v1.3","first-page":"e00014","article-title":"Expanding Role of Type II Secretion in Bacterial Pathogenesis and Beyond","volume":"85","year":"2017","journal-title":"Infect Immun"},{"key":"2023011217451315000_2023.01.10.523476v1.4","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1003117"},{"key":"2023011217451315000_2023.01.10.523476v1.5","doi-asserted-by":"publisher","DOI":"10.1038\/nsmb.3350"},{"key":"2023011217451315000_2023.01.10.523476v1.6","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01344-17"},{"key":"2023011217451315000_2023.01.10.523476v1.7","first-page":"e00521","article-title":"Structure and Membrane Topography of the Vibrio-Type Secretin Complex from the Type 2 Secretion System of Enteropathogenic Escherichia coli","volume":"200","year":"2018","journal-title":"J Bacteriol"},{"key":"2023011217451315000_2023.01.10.523476v1.8","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1038\/s41564-018-0148-0","article-title":"Structural insight into the assembly of the type II secretion system pilotin\u2013secretin complex from enterotoxigenic Escherichia coli","volume":"3","year":"2018","journal-title":"Nat Microbiol"},{"key":"2023011217451315000_2023.01.10.523476v1.9","doi-asserted-by":"publisher","DOI":"10.1101\/397794"},{"key":"2023011217451315000_2023.01.10.523476v1.10","doi-asserted-by":"crossref","first-page":"e1007731","DOI":"10.1371\/journal.ppat.1007731","article-title":"Structure and assembly of pilotin-dependent and -independent secretins of the type II secretion system","volume":"15","year":"2019","journal-title":"Plos Pathog"},{"key":"2023011217451315000_2023.01.10.523476v1.11","first-page":"e00617","article-title":"Alterations in Peptidoglycan Cross-Linking Suppress the Secretin Assembly Defect Caused by Mutation of GspA in the Type II Secretion System","volume":"199","year":"2017","journal-title":"J Bacteriol"},{"key":"2023011217451315000_2023.01.10.523476v1.12","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0102038"},{"key":"2023011217451315000_2023.01.10.523476v1.13","first-page":"e00253","article-title":"Scaffolding Protein GspB\/OutB Facilitates Assembly of the Dickeya dadantii Type 2 Secretion System by Anchoring the Outer Membrane Secretin Pore to the Inner Membrane and to the Peptidoglycan Cell Wall","volume":"13","year":"2022","journal-title":"Mbio"},{"key":"2023011217451315000_2023.01.10.523476v1.14","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.06394-11"},{"key":"2023011217451315000_2023.01.10.523476v1.15","doi-asserted-by":"crossref","first-page":"20150030","DOI":"10.1098\/rstb.2015.0030","article-title":"Outer membrane lipoprotein biogenesis: Lol is not the end","volume":"370","year":"2015","journal-title":"Philosophical Transactions Royal Soc B Biological Sci"},{"key":"2023011217451315000_2023.01.10.523476v1.16","doi-asserted-by":"publisher","DOI":"10.1128\/jb.178.12.3544-3549.1996"},{"key":"2023011217451315000_2023.01.10.523476v1.17","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/19.24.6697"},{"key":"2023011217451315000_2023.01.10.523476v1.18","first-page":"17462","article-title":"Protein secretion by gram-negative bacteria. Characterization of two membrane proteins required for pullulanase secretion by Escherichia coli K-12","volume":"264","year":"1989","journal-title":"J Biological Chem"},{"key":"2023011217451315000_2023.01.10.523476v1.19","doi-asserted-by":"crossref","first-page":"3531","DOI":"10.1002\/j.1460-2075.1987.tb02679.x","article-title":"Cloning and expression in Escherichia coli of the Klebsiella pneumoniae genes for production, surface localization and secretion of the lipoprotein pullulanase","volume":"6","year":"1987","journal-title":"Embo J"},{"key":"2023011217451315000_2023.01.10.523476v1.20","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M203740200"},{"key":"2023011217451315000_2023.01.10.523476v1.21","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01115-06"},{"key":"2023011217451315000_2023.01.10.523476v1.22","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00901-15"},{"key":"2023011217451315000_2023.01.10.523476v1.23","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-019-0591-8"},{"key":"2023011217451315000_2023.01.10.523476v1.24","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2013.06.013"},{"key":"2023011217451315000_2023.01.10.523476v1.25","doi-asserted-by":"publisher","DOI":"10.1038\/nsmb.1910"},{"key":"2023011217451315000_2023.01.10.523476v1.26","doi-asserted-by":"publisher","DOI":"10.1128\/jb.174.17.5549-5559.1992"},{"key":"2023011217451315000_2023.01.10.523476v1.27","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0805309105"},{"key":"2023011217451315000_2023.01.10.523476v1.28","doi-asserted-by":"crossref","unstructured":"Baba, T. et al. Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol Syst Biol 2, 2006.0008-2006.0008 (2006).","DOI":"10.1038\/msb4100050"},{"key":"2023011217451315000_2023.01.10.523476v1.29","doi-asserted-by":"publisher","DOI":"10.1038\/msb.2009.92"},{"key":"2023011217451315000_2023.01.10.523476v1.30","doi-asserted-by":"crossref","unstructured":"Hu, B. et al. Visualization of the type III secretion sorting platform of Shigella flexneri. Proceedings of the National Academy of Sciences 112, (2015).","DOI":"10.1073\/pnas.1411610112"},{"key":"2023011217451315000_2023.01.10.523476v1.31","doi-asserted-by":"publisher","DOI":"10.3791\/60517"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2023.01.10.523476","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T04:07:08Z","timestamp":1768450028000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2023.01.10.523476"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,10]]},"references-count":31,"URL":"https:\/\/doi.org\/10.1101\/2023.01.10.523476","relation":{},"subject":[],"published":{"date-parts":[[2023,1,10]]},"subtype":"preprint"}}