{"paper_id":"06d62e08-362e-4da6-801d-208d814adfcd","body_text":"Delineation of cold-responsive stimulation of lipolysis by hormone-sensitive lipase to increase thermogenesis in brown fat | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Delineation of cold-responsive stimulation of lipolysis by hormone-sensitive lipase to increase thermogenesis in brown fat Sheng-Cai Lin, Baoshen He, Sihan Lv, Lin Liu, Yingying Li, Zhiying Hu, and 13 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8874878/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Cold stimulates adrenergic signaling to trigger rapid lipolysis in brown adipose tissue (BAT) for adaptive thermogenesis. Protein kinase A (PKA) and hormone-sensitive lipase (HSL) are required in mediating the adrenergic signaling; however, the mechanism underlying HSL activation and translocation to lipid droplets remains unclear. Here, we demonstrate that cold-induced activation of PKA phosphorylates and activates the new substrate lysine acetyltransferase 5 (KAT5), which acetylates HSL. We further show that acetylation licenses HSL to be phosphorylated for activation by PKA. After undergoing both acetylation and phosphorylation, HSL translocates to lipid droplets through binding to the lipid droplet protein perilipin 1 (PLIN1). In vivo , BAT-specific knockout of Kat5 severely impairs cold-induced lipolysis and thermogenesis in mice; re-introduction of wild-type KAT5, but not a PKA-unphosphorylatable mutant (KAT5 S86A ), rescues this defect. Strikingly, thermogenesis is also restored by an acetylation-mimetic HSL mutant (HSL 2KQ ), but not by an acetylation-defective mutant (HSL 2KR ), reassuring the critical role of the acetylation on HSL. Together, our findings elucidate a PKA-KAT5-HSL signaling pathway, in which KAT5-mediated acetylation acts as a mandatory amplifier, enabling HSL phosphorylation and subsequent the rapid lipolysis required for robust thermogenesis. Our work hence provides a comprehensive understanding of how cold-stimulated adrenergic signaling promotes lipid mobilization to fuel heat production. Biological sciences/Physiology/Metabolism/Fat metabolism Biological sciences/Molecular biology/Post-translational modifications/Phosphorylation Biological sciences/Molecular biology/Post-translational modifications/Acetylation Full Text Additional Declarations There is NO Competing Interest. Supplementary Files Reportingsummary.pdf Reporting summary Cite Share Download PDF Status: Under Review Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-8874878\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Article\",\"associatedPublications\":[],\"authors\":[{\"id\":618402416,\"identity\":\"4aca4cfc-db32-4ea8-8f1b-32c401d7858c\",\"order_by\":0,\"name\":\"Sheng-Cai Lin\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAq0lEQVRIiWNgGAWjYBACPmYQWQHhSBClhQ2s5QxJWkAEYxtJWth5D3/4Oe9wtMEB5oO3eRjs8ohwGF+aZO+2tNwNB9iSrXkYkouJ0MJjxsy4zQaohcdMmofhQGIDEVqMPzPOkQBq4f9GtBYDacYGsC1sRGsxk+w5lpY78zCbseUcg2TCWvj5zxh/+FFzOLfvePPDG28q7AhrQQBwnBoQr34UjIJRMApGAR4AAAUGMZTekwobAAAAAElFTkSuQmCC\",\"orcid\":\"https://orcid.org/0000-0003-1993-8376\",\"institution\":\"Xiamen University\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Sheng-Cai\",\"middleName\":\"\",\"lastName\":\"Lin\",\"suffix\":\"\"},{\"id\":618402417,\"identity\":\"9cdf6c19-8ef9-4032-aa06-639cc6d6e082\",\"order_by\":1,\"name\":\"Baoshen He\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Henan university\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Baoshen\",\"middleName\":\"\",\"lastName\":\"He\",\"suffix\":\"\"},{\"id\":618402418,\"identity\":\"cbe457ef-13a4-4304-84af-2c10860fa828\",\"order_by\":2,\"name\":\"Sihan Lv\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Zhongzhou Laboratory for Integrative Biology, School of Basic Medical Sciences, Henan University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Sihan\",\"middleName\":\"\",\"lastName\":\"Lv\",\"suffix\":\"\"},{\"id\":618402419,\"identity\":\"bf73c687-6ea2-49d7-a300-22e2c96bdf94\",\"order_by\":3,\"name\":\"Lin Liu\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Zhongzhou Laboratory for Integrative Biology, School of Basic Medical Sciences, Henan University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Lin\",\"middleName\":\"\",\"lastName\":\"Liu\",\"suffix\":\"\"},{\"id\":618402420,\"identity\":\"24eaa485-67df-433d-8af1-3677e968ebe4\",\"order_by\":4,\"name\":\"Yingying Li\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Zhongzhou Laboratory for Integrative Biology, School of Basic Medical Sciences, Henan University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Yingying\",\"middleName\":\"\",\"lastName\":\"Li\",\"suffix\":\"\"},{\"id\":618402421,\"identity\":\"82dfa06b-2117-4588-a908-3d8feabd211d\",\"order_by\":5,\"name\":\"Zhiying Hu\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Zhongzhou Laboratory for Integrative Biology, School of Basic Medical Sciences, Henan University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Zhiying\",\"middleName\":\"\",\"lastName\":\"Hu\",\"suffix\":\"\"},{\"id\":618402422,\"identity\":\"d6202e3e-1262-4e56-9d3e-191010f1a714\",\"order_by\":6,\"name\":\"Bowen Zhang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Zhongzhou Laboratory for Integrative Biology, School of Basic Medical Sciences, Henan University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Bowen\",\"middleName\":\"\",\"lastName\":\"Zhang\",\"suffix\":\"\"},{\"id\":618402423,\"identity\":\"359507a2-0db7-45bb-9393-04a4dbc411de\",\"order_by\":7,\"name\":\"Yaying Wu\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Xiamen University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Yaying\",\"middleName\":\"\",\"lastName\":\"Wu\",\"suffix\":\"\"},{\"id\":618402424,\"identity\":\"cb402967-ec8d-4ae4-bd0f-377063649b4a\",\"order_by\":8,\"name\":\"Zheni Xu\",\"email\":\"\",\"orcid\":\"https://orcid.org/0000-0001-7926-0259\",\"institution\":\"Xiamen University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Zheni\",\"middleName\":\"\",\"lastName\":\"Xu\",\"suffix\":\"\"},{\"id\":618402425,\"identity\":\"f801adb3-1c5d-4b01-959f-7f34032e7948\",\"order_by\":9,\"name\":\"Cixiong Zhang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"State Key Laboratory of Cellular Stress Biology, Innovation Center for Cell Signaling Network, School of Life Sciences, Xiamen University, Xiamen, Fujian 361102\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Cixiong\",\"middleName\":\"\",\"lastName\":\"Zhang\",\"suffix\":\"\"},{\"id\":618402426,\"identity\":\"5e0d4065-f56c-40ac-9ffc-1baf8de94928\",\"order_by\":10,\"name\":\"Lei Zhang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Lei\",\"middleName\":\"\",\"lastName\":\"Zhang\",\"suffix\":\"\"},{\"id\":618402427,\"identity\":\"6016e4ec-2bb9-4a03-abb4-6638fed60008\",\"order_by\":11,\"name\":\"Xi Huang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Xiamen University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Xi\",\"middleName\":\"\",\"lastName\":\"Huang\",\"suffix\":\"\"},{\"id\":618402428,\"identity\":\"1e6a76b3-b3cd-49c0-aa7b-967a854fbf19\",\"order_by\":12,\"name\":\"Jianfeng Wu\",\"email\":\"\",\"orcid\":\"https://orcid.org/0000-0002-8992-2030\",\"institution\":\"Xiamen University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Jianfeng\",\"middleName\":\"\",\"lastName\":\"Wu\",\"suffix\":\"\"},{\"id\":618402429,\"identity\":\"e6203800-9465-46e1-bc49-8b777f7c3c99\",\"order_by\":13,\"name\":\"Su Chen\",\"email\":\"\",\"orcid\":\"https://orcid.org/0000-0001-8804-8989\",\"institution\":\"Zhongzhou Laboratory for Integrative Biology, School of Basic Medical Sciences, Henan University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Su\",\"middleName\":\"\",\"lastName\":\"Chen\",\"suffix\":\"\"},{\"id\":618402430,\"identity\":\"f5d5c43a-11ba-468c-b70b-9c911ca876f3\",\"order_by\":14,\"name\":\"Wei Hong\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Xiamen University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Wei\",\"middleName\":\"\",\"lastName\":\"Hong\",\"suffix\":\"\"},{\"id\":618402431,\"identity\":\"8f571be0-64cf-4d09-aa2a-5b173c750c12\",\"order_by\":15,\"name\":\"Tianyu Tang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Faculty of Medicine and Life Sciences, Xiamen University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Tianyu\",\"middleName\":\"\",\"lastName\":\"Tang\",\"suffix\":\"\"},{\"id\":618402432,\"identity\":\"c84b0222-98c3-4b56-a0f8-8f1cb7334fbf\",\"order_by\":16,\"name\":\"Chen-Song Zhang\",\"email\":\"\",\"orcid\":\"https://orcid.org/0000-0001-5218-0101\",\"institution\":\"Xiamen University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Chen-Song\",\"middleName\":\"\",\"lastName\":\"Zhang\",\"suffix\":\"\"},{\"id\":618402433,\"identity\":\"ce47b56e-9835-41e7-9613-c51382fe50db\",\"order_by\":17,\"name\":\"Terytty Yang Li\",\"email\":\"\",\"orcid\":\"https://orcid.org/0000-0003-3684-7935\",\"institution\":\"Fudan University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Terytty\",\"middleName\":\"Yang\",\"lastName\":\"Li\",\"suffix\":\"\"},{\"id\":618402434,\"identity\":\"6f7eadb4-c7f1-4993-b97d-3ebf593ae1cd\",\"order_by\":18,\"name\":\"Shu-Yong Lin\",\"email\":\"\",\"orcid\":\"https://orcid.org/0000-0002-4419-1713\",\"institution\":\"Xiamen University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Shu-Yong\",\"middleName\":\"\",\"lastName\":\"Lin\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2026-02-13 18:55:56\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-8874878/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-8874878/v1\",\"draftVersion\":[],\"editorialEvents\":[],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":106421510,\"identity\":\"0fe8437f-863b-4f2b-9dd1-6b5f9c739571\",\"added_by\":\"auto\",\"created_at\":\"2026-04-08 11:13:10\",\"extension\":\"pdf\",\"order_by\":1,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":40704714,\"visible\":true,\"origin\":\"\",\"legend\":\"Article File\",\"description\":\"\",\"filename\":\"PKAKAT5HSLmanuscript260213.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8874878/v1_covered_7c757235-8950-48ae-a840-fd895262aa40.pdf\"},{\"id\":106421505,\"identity\":\"5a70fe06-1c03-4418-a97f-2ed5afee2e24\",\"added_by\":\"auto\",\"created_at\":\"2026-04-08 11:12:59\",\"extension\":\"pdf\",\"order_by\":1,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":339163,\"visible\":true,\"origin\":\"\",\"legend\":\"Reporting summary\",\"description\":\"\",\"filename\":\"Reportingsummary.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-8874878/v1/a41df57bc2e0c796a6e5af13.pdf\"}],\"financialInterests\":\"There is \\u003cb\\u003eNO\\u003c/b\\u003e Competing Interest.\",\"formattedTitle\":\"Delineation of cold-responsive stimulation of lipolysis by hormone-sensitive lipase to increase thermogenesis in brown fat\",\"fulltext\":[],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":false,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":true,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":false,\"isAuthorSuppliedPdf\":true,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":true,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"nature-portfolio\",\"isNatureJournal\":true,\"hasQc\":false,\"allowDirectSubmit\":false,\"externalIdentity\":\"\",\"sideBox\":\"\",\"snPcode\":\"\",\"submissionUrl\":\"\",\"title\":\"Nature Portfolio\",\"twitterHandle\":\"\",\"acdcEnabled\":false,\"dfaEnabled\":false,\"editorialSystem\":\"ejp\",\"reportingPortfolio\":\"\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false},\"keywords\":\"\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-8874878/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-8874878/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"Cold stimulates adrenergic signaling to trigger rapid lipolysis in brown adipose tissue (BAT) for adaptive thermogenesis. Protein kinase A (PKA) and hormone-sensitive lipase (HSL) are required in mediating the adrenergic signaling; however, the mechanism underlying HSL activation and translocation to lipid droplets remains unclear. Here, we demonstrate that cold-induced activation of PKA phosphorylates and activates the new substrate lysine acetyltransferase 5 (KAT5), which acetylates HSL. We further show that acetylation licenses HSL to be phosphorylated for activation by PKA. After undergoing both acetylation and phosphorylation, HSL translocates to lipid droplets through binding to the lipid droplet protein perilipin 1 (PLIN1). \\u003ci\\u003eIn vivo\\u003c/i\\u003e, BAT-specific knockout of \\u003ci\\u003eKat5\\u003c/i\\u003e severely impairs cold-induced lipolysis and thermogenesis in mice; re-introduction of wild-type KAT5, but not a PKA-unphosphorylatable mutant (KAT5\\u003csup\\u003eS86A\\u003c/sup\\u003e), rescues this defect. Strikingly, thermogenesis is also restored by an acetylation-mimetic HSL mutant (HSL\\u003csup\\u003e2KQ\\u003c/sup\\u003e), but not by an acetylation-defective mutant (HSL\\u003csup\\u003e2KR\\u003c/sup\\u003e), reassuring the critical role of the acetylation on HSL. Together, our findings elucidate a PKA-KAT5-HSL signaling pathway, in which KAT5-mediated acetylation acts as a mandatory amplifier, enabling HSL phosphorylation and subsequent the rapid lipolysis required for robust thermogenesis. Our work hence provides a comprehensive understanding of how cold-stimulated adrenergic signaling promotes lipid mobilization to fuel heat production.\",\"manuscriptTitle\":\"Delineation of cold-responsive stimulation of lipolysis by hormone-sensitive lipase to increase thermogenesis in brown fat\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2026-04-08 11:11:54\",\"doi\":\"10.21203/rs.3.rs-8874878/v1\",\"editorialEvents\":[],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"nature-communications\",\"isNatureJournal\":true,\"hasQc\":false,\"allowDirectSubmit\":false,\"externalIdentity\":\"NCOMMS\",\"sideBox\":\"Learn more about [Nature Communications](http://www.nature.com/ncomms/)\",\"snPcode\":\"\",\"submissionUrl\":\"https://mts-ncomms.nature.com/\",\"title\":\"Nature Communications\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"ejp\",\"reportingPortfolio\":\"Nature Communications\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false}}],\"origin\":\"\",\"ownerIdentity\":\"a0c79d48-99d1-4592-a543-5c233ee26115\",\"owner\":[],\"postedDate\":\"April 8th, 2026\",\"published\":true,\"recentEditorialEvents\":[{\"type\":\"editorInvitedReview\",\"content\":\"This content is not available.\",\"date\":\"2026-05-05T12:31:53+00:00\",\"index\":4,\"fulltext\":\"This content is not available.\"},{\"type\":\"editorInvitedReview\",\"content\":\"This content is not available.\",\"date\":\"2026-05-01T06:24:04+00:00\",\"index\":3,\"fulltext\":\"This content is not available.\"}],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"under-review\",\"subjectAreas\":[{\"id\":65804173,\"name\":\"Biological sciences/Physiology/Metabolism/Fat metabolism\"},{\"id\":65804174,\"name\":\"Biological sciences/Molecular biology/Post-translational modifications/Phosphorylation\"},{\"id\":65804175,\"name\":\"Biological sciences/Molecular biology/Post-translational modifications/Acetylation\"}],\"tags\":[],\"updatedAt\":\"2026-04-08T11:11:54+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2026-04-08 11:11:54\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-8874878\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-8874878\",\"identity\":\"rs-8874878\",\"version\":[\"v1\"]},\"buildId\":\"XKTyCvWXoU3ODBz1xrDgd\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}