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Here, we present the cryo-electron microscopy structure of full-length human SLFN14, a member with antiviral activity and linked to an inherited bleeding disorder. SLFN14 is composed of an RNase domain, a SWADL domain, and a two-lobe helicase domain. SLFN14 exhibited strong RNase activity over different substrates, and the positively charged patches at the valley of the RNase domain, which contains the thrombocytopenia-related missense mutation sites, are crucial for binding oligonucleotides. SLFN14 lacks helicase activity, which can be attributed to the inability to bind ATP and the absence of positive charges at the canonical DNA-binding site of its RecA-like folds. SLFN14 is structurally similar to SLFN11, but differs from SLFN5 in the orientation of the helicase domain. Live-cell fluorescence resonance energy transfer (FRET) assays and AlphaFold2 analysis hinted that SLFN14 may adopt multiple conformations in cells. These results provide detailed structural and biochemical features of SLFN14, and greatly expand our knowledge of the functional diversity of the SLFN family.<\/jats:p>","DOI":"10.1093\/nar\/gkaf484","type":"journal-article","created":{"date-parts":[[2025,5,21]],"date-time":"2025-05-21T09:07:41Z","timestamp":1747818461000},"source":"Crossref","is-referenced-by-count":4,"title":["Structural and functional characterization of human SLFN14"],"prefix":"10.1093","volume":"53","author":[{"given":"Meng","family":"Luo","sequence":"first","affiliation":[{"name":"State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center , Guangzhou 510060 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xudong","family":"Jia","sequence":"additional","affiliation":[{"name":"State Key Lab for Biocontrol, School of Life Sciences, Sun Yat-sen University , Guangzhou 510275 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zi-Wen","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center , Guangzhou 510060 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin- Yu","family":"Yang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center , Guangzhou 510060 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wen","family":"Wang","sequence":"additional","affiliation":[{"name":"MOE Key Laboratory of Laser Life Science & Guangdong Provincial Key Laboratory of Laser Life Science, College of Biophotonics, School of Optoelectronic Science and Engineering, South China Normal University , Guangzhou 510631 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiazhen","family":"Chen","sequence":"additional","affiliation":[{"name":"Institute of Human Virology, Key Laboratoy of Tropical Disease Control of Ministry of Education, Zhongshan School of Medicine, Sun Yat-sen University , Guangzhou 510080 ,","place":["China"]},{"name":"Zhongshan School of Medicine, Sun Yat-sen University Department of Immunology, , Guangzhou 510080 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun-Ying","family":"Ou","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center , Guangzhou 510060 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian-Xiong","family":"Feng","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center , Guangzhou 510060 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bing","family":"Yu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center , Guangzhou 510060 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sheng","family":"Wang","sequence":"additional","affiliation":[{"name":"Shanghai Zelixir Biotech Company Ltd , Shanghai 200030 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2121-365X","authenticated-orcid":false,"given":"Lin","family":"Huang","sequence":"additional","affiliation":[{"name":"Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Guangdong-Hong Kong Joint Laboratory for RNA Medicine, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University , Guangzhou 510120 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Neil V","family":"Morgan","sequence":"additional","affiliation":[{"name":"School of Medical Sciences, College of Medicine and Health, University of Birmingham Department of Cardiovascular Sciences, , Edgbaston B15 2TT ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9973-8130","authenticated-orcid":false,"given":"Kai","family":"Deng","sequence":"additional","affiliation":[{"name":"Institute of Human Virology, Key Laboratoy of Tropical Disease Control of Ministry of Education, Zhongshan School of Medicine, Sun Yat-sen University , Guangzhou 510080 ,","place":["China"]},{"name":"Zhongshan School of Medicine, Sun Yat-sen University Department of Immunology, , Guangzhou 510080 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Cancer Center , Guangzhou 510060 ,","place":["China"]},{"name":"Integrated Traditional Chinese and Western Medicine Research Center, Sun Yat-sen University Cancer Center , Guangzhou 510060 ,","place":["China"]},{"name":"Sun Yat-sen University Cancer Center Gansu Hospital Department of Experimental Research, , Lanzhou 730000 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2025,6,4]]},"reference":[{"key":"2025061005174010000_B1","doi-asserted-by":"publisher","first-page":"2238","DOI":"10.3390\/cells10092238","article-title":"Schlafens: emerging proteins in cancer cell biology","volume":"10","author":"Al-Marsoummi","year":"2021","journal-title":"Cells"},{"key":"2025061005174010000_B2","doi-asserted-by":"publisher","DOI":"10.3390\/v14020442","article-title":"Schlafens can put viruses to 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