{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T13:29:00Z","timestamp":1788960540181,"version":"build-2803163510"},"reference-count":42,"publisher":"Oxford University Press (OUP)","issue":"20","license":[{"start":{"date-parts":[[2025,11,8]],"date-time":"2025-11-08T00:00:00Z","timestamp":1762560000000},"content-version":"vor","delay-in-days":11,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32300036 to H. Y."],"award-info":[{"award-number":["32300036 to H. Y."]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Foundation of Tianjin Science and Technology Commission","award":["23ZYCGSY00750"],"award-info":[{"award-number":["23ZYCGSY00750"]}]},{"DOI":"10.13039\/501100010229","name":"Natural Science Foundation of Tianjin Municipal Science and Technology Commission","doi-asserted-by":"publisher","award":["23JCZDJC00410"],"award-info":[{"award-number":["23JCZDJC00410"]}],"id":[{"id":"10.13039\/501100010229","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,10,28]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Defense-associated reverse transcriptase (DRT) systems play a crucial role in prokaryotic defense mechanisms against phage infections. Among the DRT family, DRT2, DRT3, and DRT9 systems employ protein\u2013noncoding RNA (ncRNA) co-regulatory mechanisms to execute defense functions. Here, we focus on the DRT2 system from Klebsiella pneumoniae, which consists of a reverse transcriptase (RT) and an essential ncRNA component. Using biochemical and structural approaches, we determine the structure of the DRT2 system and reveal detailed interaction modes between the DRT2-RT protein and the ncRNA, especially mediated by specialized anchoring loops and pseudoknot-related structures. The RT protein adopts a conventional \u201cright-hand\u201d fold, while a flexible region of the ncRNA exhibits dynamic conformations, likely serving as the template for reverse transcription. DRT2 mediates reverse transcription through a conserved DDD catalytic triad that coordinates a divalent Mg\u00b2\u207a ion. Notably, a short DNA primer-ncRNA duplex is accommodated in a positively charged pocket formed by the thumb and fingers domains, and both interaction analysis and mutagenesis studies confirm that duplex stabilization is essential for activity. Structural comparison and phylogenetic studies of DRT2 and other RT proteins, such as group II introns and UG\/Abi RTs, highlight the unique adaptation with a straight extended thumb domain and specialized structures for ncRNA-binding, exemplifying an evolutionary trajectory of RT proteins. In conclusion, our findings expand the understanding of the distinctive characteristics of the DRT2 system and the diversity of prokaryotic antiviral strategies.<\/jats:p>","DOI":"10.1093\/nar\/gkaf1135","type":"journal-article","created":{"date-parts":[[2025,11,9]],"date-time":"2025-11-09T11:26:38Z","timestamp":1762687598000},"source":"Crossref","is-referenced-by-count":1,"title":["Molecular mechanism of the type 2 defense-associated reverse transcriptase"],"prefix":"10.1093","volume":"53","author":[{"given":"Zhikun","family":"Liu","sequence":"first","affiliation":[{"name":"Tianjin Medical University Cancer Institute and Hospital , State Key Laboratory of Experimental Hematology, Key Laboratory of Immune Microenvironment and Disease (Ministry of Education), The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, National Clinical Research Center for Cancer, Tianjin Institute of Immunology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070 ,","place":["China"]},{"name":"Tianjin Key Laboratory of Cellular Homeostasis and Disease, Tianjin\u2019s Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy , Department of Biochemistry and Molecular Biology, Tianjin Medical University, Tianjin 300070 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fumeng","family":"Liao","sequence":"additional","affiliation":[{"name":"Tianjin Medical University Cancer Institute and Hospital , State Key Laboratory of Experimental Hematology, Key Laboratory of Immune Microenvironment and Disease (Ministry of Education), The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, National Clinical Research Center for Cancer, Tianjin Institute of Immunology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070 ,","place":["China"]},{"name":"Tianjin Key Laboratory of Cellular Homeostasis and Disease, Tianjin\u2019s Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy , Department of Biochemistry and Molecular Biology, Tianjin Medical University, Tianjin 300070 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenqi","family":"Wu","sequence":"additional","affiliation":[{"name":"Tianjin Medical University Cancer Institute and Hospital , State Key Laboratory of Experimental Hematology, Key Laboratory of Immune Microenvironment and Disease (Ministry of Education), The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, National Clinical Research Center for Cancer, Tianjin Institute of Immunology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070 ,","place":["China"]},{"name":"Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin\u2019s Clinical Research Center for Cancer Department of Gastric Surgery, , Tianjin 300060 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chendi","family":"Zhang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University , Wuhan 430062 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caidie","family":"Yue","sequence":"additional","affiliation":[{"name":"Tianjin Medical University Cancer Institute and Hospital, State Key Laboratory of Experimental Hematology, Key Laboratory of Immune Microenvironment and Disease (Ministry of Education), The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, National Clinical Research Center for Cancer, Tianjin Institute of Immunology, School of Basic Medical Sciences, Tianjin Medical University , Tianjin 300070 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Innovation Center for Medical Epigenetics, National Clinical Research Center for Cancer, Tianjin Institute of Immunology, School of Basic Medical Sciences, Tianjin Medical University , Tianjin 300070 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingcan","family":"Liu","sequence":"additional","affiliation":[{"name":"Tianjin Medical University Cancer Institute and Hospital, State Key Laboratory of Experimental Hematology, Key Laboratory of Immune Microenvironment and Disease (Ministry of Education), The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, National Clinical Research Center for Cancer, Tianjin Institute of Immunology, School of Basic Medical Sciences, Tianjin Medical University , Tianjin 300070 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bin","family":"Liu","sequence":"additional","affiliation":[{"name":"Tianjin Medical University Cancer Institute and Hospital, State Key Laboratory of Experimental Hematology, Key Laboratory of Immune Microenvironment and Disease (Ministry of Education), The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, National Clinical Research Center for Cancer, Tianjin Institute of Immunology, School of Basic Medical Sciences, Tianjin Medical University , Tianjin 300070 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Han","sequence":"additional","affiliation":[{"name":"School of Basic Medical Sciences, Tianjin Medical University Department of Anatomy, , Tianjin 300070 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chuyun","family":"Zhang","sequence":"additional","affiliation":[{"name":"Tianjin Medical University Cancer Institute and Hospital, State Key Laboratory of Experimental Hematology, Key Laboratory of Immune Microenvironment and Disease (Ministry of Education), The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, National Clinical Research Center for Cancer, Tianjin Institute of Immunology, School of Basic Medical Sciences, Tianjin Medical University , Tianjin 300070 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoshen","family":"Wang","sequence":"additional","affiliation":[{"name":"Tianjin Medical University Cancer Institute and Hospital, State Key Laboratory of Experimental Hematology, Key Laboratory of Immune Microenvironment and Disease (Ministry of Education), The Province and Ministry Co-sponsored Collaborative Innovation Center for Medical Epigenetics, National Clinical Research Center for Cancer, Tianjin Institute of Immunology, School of Basic Medical Sciences, Tianjin Medical University , Tianjin 300070 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