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Sci.","Veterinary Sciences"],"abstract":"<jats:p>Regulation of Duck Tembusu virus (DTMUV) non-structural protein 1 (NS1) remains unclear. Exogenous and endogenous co-immunoprecipitation (Co-IP) assays demonstrated an association between the T-complex protein 1 subunit 2 (CCT2) and NS1. Further experiments mapped the NS1 region required for assembly of this complex to amino acid residues 172-352 of NS1.DTMUV increased CCT2 expression; CCT2 overexpression increased NS1 accumulation, whereas CCT2 knockdown delayed NS1 and envelope (E) protein accumulation. Although NS1 also associated with chaperonin-containing T-complex protein 1 subunit 6 (CCT6), CCT6 knockdown did not significantly alter NS1 messenger RNA (mRNA) or clearly change NS1 protein abundance. NS1 underwent lysine 48 (K48)- and lysine 63 (K63)-linked polyubiquitination. Increasing tripartite motif-containing protein 21 (TRIM21) reduced NS1 abundance and enhanced NS1-associated ubiquitin signals; the proteasome inhibitor MG132 promoted NS1 accumulation, which was highest with CCT2 co-expression, supporting proteasome-related turnover and CCT2-associated stabilization without establishing direct TRIM21 ubiquitination. CCT2 interacted with microtubule-associated protein 1 light chain 3C (LC3C), and DTMUV infection was associated with reduced late-stage lipidated LC3 (LC3-II) abundance and SQSTM1 mRNA expression. In dose-gradient Co-IP experiments, increasing NS1 expression was associated with a dose-associated reduction in CCT2 recovery in LC3C immunoprecipitates; the concomitant reduction in immunoprecipitated LC3C across the NS1 dose gradient precludes conclusions regarding direct competition or a binding capacity-dependent mechanism. These findings indicate that CCT2 is a DTMUV-associated host factor capable of stabilizing intracellular NS1 protein and promoting the production of infectious DTMUV progeny. CCT2 contributes to the homeostasis of NS1 protein.<\/jats:p>","DOI":"10.3390\/vetsci13090979","type":"journal-article","created":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T17:56:41Z","timestamp":1790099801000},"page":"979","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["NS1 of Duck Tembusu Virus Interacts with CCT2: Reduced CCT2 Recovery in LC3C Co-Immunoprecipitation In Vitro and the Proviral Role of CCT2 in DTMUV Infection"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-5907-2113","authenticated-orcid":false,"given":"Zekun","family":"Yang","sequence":"first","affiliation":[{"name":"Zhaoqing Branch of Guangdong Laboratory of Lingnan Modern Agricultural Science and Technology, Zhaoqing 526238, 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