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To establish an intracellular niche conducive to replication,\n                  <jats:italic>L. pneumophila<\/jats:italic>\n                  translocates a diverse array of effector proteins that manipulate various host cellular processes, including the actin cytoskeleton. In a screen for effectors that alter actin dynamics, we identified a\n                  <jats:italic>Legionella<\/jats:italic>\n                  effector, Lfat1 (lpg1387), which colocalizes with the actin cytoskeleton in eukaryotic cells. Lfat1 specifically binds F-actin through a novel actin-binding domain (ABD). High-resolution cryo-electron microscopy (cryoEM) analysis revealed that this ABD forms a long \u03b1-helix hairpin, with its tip interacting with subdomains I and II of two adjacent actin molecules within the F-actin filament. Interestingly, while individual \u03b1-helices of the hairpin fail to bind F-actin, co-expression as separate fusion proteins restores binding activity. Furthermore, we demonstrated that Lfat1 exhibits lysine fatty-acyltransferase (KFAT) activity, targeting host small GTPases. These findings establish a foundation for studying the KFAT family of bacterial toxins and uncover a novel F-actin binding motif, providing an alternative F-actin marker with notable flexibility.\n                <\/jats:p>","DOI":"10.1101\/2025.04.18.649563","type":"posted-content","created":{"date-parts":[[2025,4,21]],"date-time":"2025-04-21T09:45:11Z","timestamp":1745228711000},"source":"Crossref","is-referenced-by-count":0,"title":["Cryo-EM structure revealed a novel F-actin binding motif in a\n                  <i>Legionella pneumophila<\/i>\n                  lysine fatty-acyltransferase"],"prefix":"10.64898","author":[{"given":"Wenjie W.","family":"Zeng","sequence":"first","affiliation":[{"name":"Weill Institute for Cell and Molecular Biology, Cornell University, Ithaca, NY 14853, USA"},{"name":"Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Garrison","family":"Komaniecki","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA"},{"name":"Chemistry Department, University of Wisconsin \u2013 Parkside, Kenosha, WI 53406, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiaze","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA"},{"name":"Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06520, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hening","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA"},{"name":"Howard Hughes Medical Institute; Department of Medicine and Department of Chemistry, The University of Chicago, Chicago, IL 60637, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5064-1397","authenticated-orcid":false,"given":"Yuxin","family":"Mao","sequence":"additional","affiliation":[{"name":"Weill Institute for Cell and Molecular Biology, Cornell University, Ithaca, NY 14853, USA"},{"name":"Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026041204390715000_2025.04.18.649563v2.1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444909052925"},{"key":"2026041204390715000_2025.04.18.649563v2.2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-4658.2011.08113.x"},{"key":"2026041204390715000_2025.04.18.649563v2.3","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(80)81292-0"},{"key":"2026041204390715000_2025.04.18.649563v2.4","doi-asserted-by":"crossref","unstructured":"Barrie KR , Rebowski G , Dominguez R (2024) Mechanism of actin filament severing and capping by gelsolin. 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