{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,4,11]],"date-time":"2026-04-11T04:45:40Z","timestamp":1775882740917,"version":"3.50.1"},"posted":{"date-parts":[[2025,1,8]]},"group-title":"Microbiology","reference-count":66,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,1,8]],"date-time":"2025-01-08T00:00:00Z","timestamp":1736294400000},"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":[[2025,1,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  How tick-borne pathogens interact with their hosts has been primarily studied in vertebrates where disease is observed. Comparatively less is known about pathogen interactions within the tick. Here, we report that\n                  <jats:italic>Ixodes scapularis<\/jats:italic>\n                  ticks infected with either\n                  <jats:italic>Anaplasma phagocytophilum<\/jats:italic>\n                  (causative agent of anaplasmosis) or\n                  <jats:italic>Borrelia burgdorferi<\/jats:italic>\n                  (causative agent of Lyme disease) show activation of the ATF6 branch of the unfolded protein response (UPR). Disabling ATF6 functionally restricts pathogen survival in ticks. When stimulated, ATF6 functions as a transcription factor, but is the least understood out of the three UPR pathways. To interrogate the\n                  <jats:italic>Ixodes<\/jats:italic>\n                  ATF6 transcriptional network, we developed a custom R script to query tick promoter sequences. This revealed\n                  <jats:italic>stomatin<\/jats:italic>\n                  as a potential gene target, which has roles in lipid homeostasis and vesical transport.\n                  <jats:italic>Ixodes stomatin<\/jats:italic>\n                  was experimentally validated as a bona fide ATF6-regulated gene through luciferase reporter assays, pharmacological activators, and RNAi transcriptional repression. Silencing\n                  <jats:italic>stomatin<\/jats:italic>\n                  decreased\n                  <jats:italic>A. phagocytophilum<\/jats:italic>\n                  colonization in\n                  <jats:italic>Ixodes<\/jats:italic>\n                  and disrupted cholesterol dynamics in tick cells. Furthermore, blocking\n                  <jats:italic>stomatin<\/jats:italic>\n                  restricted cholesterol availability to the bacterium, thereby inhibiting growth and survival. Taken together, we have identified the\n                  <jats:italic>Ixodes<\/jats:italic>\n                  ATF6 pathway as a novel contributor to vector competence through Stomatin-regulated cholesterol homeostasis. Moreover, our custom, web-based transcription factor binding site search tool \u201cArthroQuest\u201d revealed that the ATF6-regulated nature of\n                  <jats:italic>stomatin<\/jats:italic>\n                  is unique to blood-feeding arthropods. Collectively, these findings highlight the importance of studying fundamental processes in non-model organisms.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>IMPORTANCE<\/jats:title>\n                  <jats:p>\n                    Host-pathogen interactions for tick-borne pathogens like\n                    <jats:italic>Anaplasma phagocytophilum<\/jats:italic>\n                    (causative agent of Anaplasmosis) have been primarily studied in mammalian hosts. Comparatively less is known about interactions within the tick. Herein, we find that tick-borne pathogens activate the cellular stress response receptor, ATF6, in\n                    <jats:italic>Ixodes<\/jats:italic>\n                    ticks. Upon activation, ATF6 is cleaved and the cytosolic portion translocates to the nucleus to function as a transcription factor that coordinates gene expression networks. Using a custom script in R to query the\n                    <jats:italic>Ixodes<\/jats:italic>\n                    ATF6 regulome,\n                    <jats:italic>stomatin<\/jats:italic>\n                    was identified as an ATF6-regulated target that supports\n                    <jats:italic>Anaplasma<\/jats:italic>\n                    colonization by facilitating cholesterol availability to the bacterium. Moreover, our custom, web-based tool \u201cArthroQuest\u201d found that the ATF6-regulated nature of\n                    <jats:italic>stomatin<\/jats:italic>\n                    is unique to arthropods. Given that lipid hijacking is common among arthropod-borne microbes, ATF6-mediated induction of\n                    <jats:italic>stomatin<\/jats:italic>\n                    may be a mechanism that is exploited in many vector-pathogen relationships for the survival and persistence of transmissible microbes. Collectively, this study identified a novel contributor to vector competence and highlights the importance of studying molecular networks in non-model organisms.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.01.08.632023","type":"posted-content","created":{"date-parts":[[2025,1,9]],"date-time":"2025-01-09T03:45:18Z","timestamp":1736394318000},"source":"Crossref","is-referenced-by-count":1,"title":["ATF6 enables pathogen infection in ticks by inducing\n                  <i>stomatin<\/i>\n                  and altering cholesterol dynamics"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2865-7031","authenticated-orcid":false,"given":"Kaylee A.","family":"Vosbigian","sequence":"first","affiliation":[{"name":"Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sarah J.","family":"Wright","sequence":"additional","affiliation":[{"name":"Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kristin L.","family":"Rosche","sequence":"additional","affiliation":[{"name":"Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elis A.","family":"Fisk","sequence":"additional","affiliation":[{"name":"Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elisabeth","family":"Ramirez-Zepp","sequence":"additional","affiliation":[{"name":"Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eric A.","family":"Shelden","sequence":"additional","affiliation":[{"name":"School of Molecular Biosciences, Washington State University, Pullman, Washington, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5212-2494","authenticated-orcid":false,"given":"Dana K.","family":"Shaw","sequence":"additional","affiliation":[{"name":"Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2026041020552907000_2025.01.08.632023v1.1","unstructured":"CDC. 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