{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T22:45:20Z","timestamp":1768517120695,"version":"3.49.0"},"posted":{"date-parts":[[2024,11,15]]},"group-title":"Microbiology","reference-count":27,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2024,11,15]],"date-time":"2024-11-15T00:00:00Z","timestamp":1731628800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,11,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  The virulence plasmid of the obligate intracellular bacterium\n                  <jats:italic>Chlamydia<\/jats:italic>\n                  encodes eight proteins. Among these, Pgp3 is crucial for pathogenicity, and Pgp4 functions as a transcriptional regulator of both plasmid and chromosomal genes. The remaining proteins, Pgp1, Pgp5, Pgp6, Pgp7, and Pgp8, are predicted to play various roles in plasmid replication or maintenance based on their amino acid sequences. However, the function of Pgp2 remains unknown, even though it is required for transformation. In this study, we utilized AlphaFold to predict the 3-dimensional (3-D) structure of\n                  <jats:italic>C. trachomatis<\/jats:italic>\n                  Pgp2. Despite a lack of apparent sequence homology, the AlphaFold structure exhibited high similarity to experimentally determined structures of several plasmid replication initiators. Notably, Pgp2 features a unique \u03b2-hairpin motif near the DNA-binding domain, absent in other plasmid replication initiators with overall 3-D structures similar to Pgp2. This \u03b2-hairpin motif was also present in AlphaFold models of Pgp2s across all 13\n                  <jats:italic>Chlamydia<\/jats:italic>\n                  species. To assess its significance, we engineered a plasmid lacking the 11 amino acids constituting the \u03b2-hairpin motif in\n                  <jats:italic>C. trachomatis<\/jats:italic>\n                  Pgp2. Although this deletion did not alter the overall structure of Pgp2, the mutated plasmid failed to transform plasmid-free\n                  <jats:italic>C. trachomatis<\/jats:italic>\n                  . These findings reveal that Pgp2 is a plasmid replication initiator, with the \u03b2-hairpin motif playing a critical role in binding to its cognate iteron sequences in the replication origin of the chlamydial plasmid.\n                <\/jats:p>","DOI":"10.1101\/2024.11.14.623704","type":"posted-content","created":{"date-parts":[[2024,11,15]],"date-time":"2024-11-15T22:55:13Z","timestamp":1731711313000},"source":"Crossref","is-referenced-by-count":0,"title":["<i>Chlamydia<\/i>\n                  plasmid-encoded protein Pgp2 is a replication initiator with a unique \u03b2-hairpin necessary for iteron-binding and plasmid replication"],"prefix":"10.64898","author":[{"given":"Danny","family":"Wan","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matthew","family":"Pan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7053-5009","authenticated-orcid":false,"given":"Guangming","family":"Zhong","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4903-926X","authenticated-orcid":false,"given":"Huizhou","family":"Fan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2024111911410479000_2024.11.14.623704v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.syapm.2014.12.004"},{"key":"2024111911410479000_2024.11.14.623704v1.2","unstructured":"WHO. 2021. Global progress report on HIV, viral hepatitis and sexually transmitted infections, 2021."},{"key":"2024111911410479000_2024.11.14.623704v1.3","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0007835"},{"key":"2024111911410479000_2024.11.14.623704v1.4","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.28658-0"},{"key":"2024111911410479000_2024.11.14.623704v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2016.09.006"},{"key":"2024111911410479000_2024.11.14.623704v1.6","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20111266"},{"key":"2024111911410479000_2024.11.14.623704v1.7","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01377-07"},{"key":"2024111911410479000_2024.11.14.623704v1.8","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2019.00013"},{"key":"2024111911410479000_2024.11.14.623704v1.9","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.02576-14"},{"key":"2024111911410479000_2024.11.14.623704v1.10","doi-asserted-by":"publisher","DOI":"10.1128\/iai.00429-17"},{"key":"2024111911410479000_2024.11.14.623704v1.11","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00102-08"},{"key":"2024111911410479000_2024.11.14.623704v1.12","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01305-12"},{"key":"2024111911410479000_2024.11.14.623704v1.13","doi-asserted-by":"publisher","DOI":"10.1111\/2049-632X.12104"},{"key":"2024111911410479000_2024.11.14.623704v1.14","first-page":"4053","article-title":"Analysis of the entire nucleotide sequence of the cryptic plasmid of Chlamydia trachomatis serovar L1","volume":"16","year":"1988","journal-title":"Evidence for involvement in DNA replication. 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