{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,5,30]],"date-time":"2026-05-30T08:01:55Z","timestamp":1780128115217,"version":"3.54.0"},"posted":{"date-parts":[[2025,10,9]]},"group-title":"Molecular Biology","reference-count":58,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T00:00:00Z","timestamp":1759968000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH","award":["U19AI171110"],"award-info":[{"award-number":["U19AI171110"]}]},{"name":"French Government\u2019s Investissement d\u2019Avenir program","award":["ANR-10-LABX-62-IBEID"],"award-info":[{"award-number":["ANR-10-LABX-62-IBEID"]}]},{"name":"Fondation iXcore - iXlife - iXblue Pour La Recherche"},{"name":"Explore Donation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,27]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Viral macrodomains are promising antiviral targets that counteract host ADP-ribosylation-mediated antiviral responses in mammals. However, their role in dual-host viruses within the mosquito vector remains largely unknown. Here, we investigated the role of the Chikungunya virus (CHIKV) nsP3 macrodomain catalytic activity by mutating the active-site asparagine 24 (N24). N24 mutant viruses rapidly selected for compensatory second-site mutations at aspartate 31 (D31) in mammalian Vero cells, while interferon-competent human cells favored reversion at position 24 instead, suggesting adaptive responses to macrodomain functional impairment. Catalytically inactive double-mutant viruses replicated poorly in interferon-competent human cells but comparably to wild-type in mosquito cells. In\n                  <jats:italic>Aedes<\/jats:italic>\n                  mosquitoes\n                  <jats:italic>in vivo<\/jats:italic>\n                  , macrodomain mutations modulated CHIKV replication and infectivity in a mutation-specific manner: one mutant reduced them while another enhanced them. Biochemical and structural analyses confirmed that these phenotypes arise in the context of a catalytically inactive macrodomain that retains ADP-ribose binding capacity, with D31 mutations lining the substrate exit path. These findings demonstrate that macrodomain catalytic activity modulates CHIKV dissemination and transmission potential in\n                  <jats:italic>Aedes<\/jats:italic>\n                  mosquitoes, revealing fundamental differences in host-dependent evolutionary advantage for a catalytically inactive viral enzyme.\n                <\/jats:p>","DOI":"10.1101\/2025.10.09.681498","type":"posted-content","created":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T03:25:13Z","timestamp":1760066713000},"source":"Crossref","is-referenced-by-count":0,"title":["Macrodomain catalytic activity modulates Chikungunya virus dissemination and transmission potential in\n                  <i>Aedes<\/i>\n                  mosquitoes"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3255-0923","authenticated-orcid":false,"given":"Eugenia S.","family":"Bardossy","sequence":"first","affiliation":[{"name":"Viruses and RNAi Unit, Institut Pasteur, Universit\u00e9 Paris Cit\u00e9, F-75015 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lena","family":"Bergmann","sequence":"additional","affiliation":[{"name":"Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, CA 94158, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Annabelle","family":"Henrion-Lacritick","sequence":"additional","affiliation":[{"name":"Viruses and RNAi Unit, Institut Pasteur, Universit\u00e9 Paris Cit\u00e9, F-75015 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jared","family":"Nigg","sequence":"additional","affiliation":[{"name":"Viruses and RNAi Unit, Institut Pasteur, Universit\u00e9 Paris Cit\u00e9, F-75015 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Galen J.","family":"Correy","sequence":"additional","affiliation":[{"name":"Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, CA 94158, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alan","family":"Ashworth","sequence":"additional","affiliation":[{"name":"Helen Diller Family Comprehensive Cancer Center, University of California San Francisco, San Francisco, California, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5080-2859","authenticated-orcid":false,"given":"James S.","family":"Fraser","sequence":"additional","affiliation":[{"name":"Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, CA 94158, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria-Carla","family":"Saleh","sequence":"additional","affiliation":[{"name":"Viruses and RNAi Unit, Institut Pasteur, Universit\u00e9 Paris Cit\u00e9, F-75015 Paris, 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