{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,15]],"date-time":"2026-05-15T13:04:34Z","timestamp":1778850274264,"version":"3.51.4"},"reference-count":63,"publisher":"American Society for Microbiology","issue":"19","license":[{"start":{"date-parts":[[2015,10,1]],"date-time":"2015-10-01T00:00:00Z","timestamp":1443657600000},"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":[[2015,10]]},"abstract":"ABSTRACT<\/jats:title>\n \n A rising theme among intracellular microbes is the delivery of ankyrin repeat-containing effectors (Anks) that interact with target proteins to co-opt host cell functions.\n Orientia tsutsugamushi<\/jats:named-content>\n , an obligate intracellular bacterium and the etiologic agent of scrub typhus, encodes one of the largest Ank repertoires of any sequenced microorganism. They have been previously identified as type 1 secretion system substrates. Here,\n in silico<\/jats:italic>\n and manual sequence analyses revealed that a large proportion of\n O. tsutsugamushi<\/jats:named-content>\n strain Ikeda Anks bear a eukaryotic\/poxvirus-like F-box motif, which is known to recruit host cell SCF1 ubiquitin ligase machinery. We assessed the Anks for the ability to serve as F-box proteins. Coimmunoprecipitation assays demonstrated that F-box-containing Anks interact with overexpressed and\/or endogenous SCF1 components. When coexpressed with FLAG-Ank4_01 or FLAG-Ank9, a glutathione\n S<\/jats:italic>\n -transferase (GST)-tagged version of the SCF1 component SKP1 localized to subcellular sites of FLAG-Ank accumulation. The abilities of recombinant Anks to interact and colocalize with SKP1 were F-box dependent. GST-SKP1 precipitated\n O. tsutsugamushi<\/jats:named-content>\n -derived Ank9 from infected host cells, verifying both that the pathogen expresses Ank9 during infection and the protein's capability to bind SKP1. Aligning\n O. tsutsugamushi<\/jats:named-content>\n , poxviral, and eukaryotic F-box sequences delineated three F-box residues that are highly conserved and likely to be functionally important. Substitution of these residues ablated the ability of GFP-Ank9 to interact with GST-SKP1. These results demonstrate that\n O. tsutsugamushi<\/jats:named-content>\n strain Ikeda Anks can co-opt host cell polyubiquitination machinery, provide the first evidence that an\n O. tsutsugamushi<\/jats:named-content>\n Ank does so during infection, and advance overall understanding of microbial F-box proteins.\n <\/jats:p>\n \n IMPORTANCE<\/jats:bold>\n Ankyrin repeat-containing proteins (Anks) are important virulence factors of intracellular bacteria that mediate protein-protein interactions with host cell targets.\n Orientia tsutsugamushi<\/jats:named-content>\n , which causes a debilitating infection called scrub typhus in one of the most densely populated regions of the world, encodes one of the largest Ank armamentariums of any sequenced bacterium. This study demonstrates that\n O. tsutsugamushi<\/jats:named-content>\n strain Ikeda Anks also bear F-box motifs that interact with host cell polyubiquitination machinery. By proving that an\n Orientia<\/jats:named-content>\n -derived Ank interacts with SKP1 in infected cells, this evidences the first bona fide\n Orientia<\/jats:named-content>\n effector and the first example of an endogenous F-box-containing Ank\u2013mammalian-host ligand interaction for any intracellular bacterium. Also, importantly, this work identifies key residues that are essential for microbial F-box function.\n <\/jats:p>","DOI":"10.1128\/jb.00276-15","type":"journal-article","created":{"date-parts":[[2015,7,14]],"date-time":"2015-07-14T02:30:09Z","timestamp":1436841009000},"page":"3097-3109","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":37,"title":["Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs"],"prefix":"10.1128","volume":"197","author":[{"given":"Andrea R.","family":"Beyer","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lauren","family":"VieBrock","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kyle G.","family":"Rodino","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel P.","family":"Miller","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brittney K.","family":"Tegels","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Richard T.","family":"Marconi","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jason A.","family":"Carlyon","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0001466"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.4161\/hv.3.3.4009"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.micinf.2011.03.003"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/modpathol.3880385"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1086\/339908"},{"key":"e_1_3_2_7_2","doi-asserted-by":"crossref","first-page":"91","DOI":"10.4269\/ajtmh.1997.57.91","article-title":"Seroepidemiologic evidence for murine and scrub typhus in Malang, Indonesia","volume":"57","author":"Richards AL","year":"1997","unstructured":"Richards AL, Soeatmadji DW, Widodo MA, Sardjono TW, Yanuwiadi B, Hernowati TE, Baskoro AD, Roebiyoso Hakim L, Soendoro M, Rahardjo E, Putri MP, Saragih JM, Strickman D, Kelly DJ, Dasch GA, Olson JG, Church CJ, Corwin AL. 1997. Seroepidemiologic evidence for murine and scrub typhus in Malang, Indonesia. Am J Trop Med Hyg 57:91\u201395.","journal-title":"Am J Trop Med Hyg"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/149.3.330"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1086\/596576"},{"key":"e_1_3_2_10_2","doi-asserted-by":"crossref","first-page":"481","DOI":"10.4269\/ajtmh.1945.s1-25.481","article-title":"Scrub-typhus","volume":"25","author":"Browning JS","year":"1945","unstructured":"Browning JS, Raphael M, Kline EF, Coblenz A. 1945. Scrub-typhus. Am J Trop Med Hyg 25:481\u2013492.","journal-title":"Am J Trop Med Hyg"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.2307\/3273264"},{"key":"e_1_3_2_12_2","doi-asserted-by":"crossref","first-page":"26","DOI":"10.7326\/0003-4819-79-1-26","article-title":"Scrub typhus in South Vietnam. A study of 87 cases","volume":"79","author":"Berman SJ","year":"1973","unstructured":"Berman SJ, Kundin WD. 1973. 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