{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T16:56:28Z","timestamp":1791046588994,"version":"4.1.0"},"posted":{"date-parts":[[2026,9,29]]},"group-title":"Cell Biology","reference-count":29,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T00:00:00Z","timestamp":1790640000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"award":["R01HL172813"],"award-info":[{"award-number":["R01HL172813"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,29]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>\n                    <jats:italic>KCNH2<\/jats:italic>\n                    encodes Kv11.1 channel proteins that conduct the rapidly activating delayed-rectifier K+ current (I\n                    <jats:sub>Kr<\/jats:sub>\n                    ), which is critical for cardiac repolarization.\n                    <jats:italic>KCNH2<\/jats:italic>\n                    encodes two functional isoforms, Kv11.1a and Kv11.1b, via alternative transcription start sites. Kv11.1a is the principal determinant of cardiac I\n                    <jats:sub>Kr<\/jats:sub>\n                    and ventricular repolarization. The circadian clock, a transcriptional-translational feedback loop that cycles with a period of \u223c24 hours and drives the circadian expression of many genes, including\n                    <jats:italic>Kcnh2<\/jats:italic>\n                    in the mouse heart. Because daily body temperature rhythms provide a systemic signal that synchronizes cardiac circadian clocks, we tested whether physiological temperature cycles drive the circadian promoter activity of the cloned human\n                    <jats:italic>KCNH2<\/jats:italic>\n                    (h\n                    <jats:italic>KCNH2<\/jats:italic>\n                    ) promoter.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Hypothesis<\/jats:title>\n                  <jats:p>\n                    h\n                    <jats:italic>KCNH2<\/jats:italic>\n                    is a direct transcriptional target of the circadian clock, with temperature driving its promoter activity through BMAL1:CLOCK acting at a conserved tandem E-box.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>\n                    We cloned the conserved proximal promoter of\n                    <jats:italic>KCNH2<\/jats:italic>\n                    (-1631 bp upstream of Kv11.1a exon 1) to generate h\n                    <jats:italic>KCNH2<\/jats:italic>\n                    promoter luciferase reporter constructs. Constructs were transfected into C2C12 myotubes and synchronized by serum shock (static 37\u00b0C) or temperature cycling (36.5\u201338.5\u00b0C). Bioluminescence was recorded and assessed for period, phase, and amplitude. BMAL1:CLOCK dependence was tested via dominant-negative CLOCK\u039419 co-expression.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    Temperature cycling did not exhibit the rapid damping characteristic of serum-shock-synchronized oscillations, consistent with continuous entrainment by an external zeitgeber rather than a free-running oscillator. Deletion analysis identified a conserved tandem E-box required for oscillation under both serum shock and temperature cycling, and for BMAL1:CLOCK-dependent transactivation (1.75 \u00b1 0.21 vs. 0.86 \u00b1 0.06 RLU, p = 0.0038). CLOCK\u039419 reduced h\n                    <jats:italic>KCNH2<\/jats:italic>\n                    promoter amplitude under temperature cycling without altering period.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>\n                    The circadian clock regulates\n                    <jats:italic>KCNH2<\/jats:italic>\n                    promoter activity through daily temperature rhythms.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.09.28.751769","type":"posted-content","created":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T15:55:14Z","timestamp":1790697314000},"source":"Crossref","is-referenced-by-count":0,"title":["The circadian clock regulates\n                  <i>KCNH2<\/i>\n                  (hERG) promoter activity through daily temperature rhythms"],"prefix":"10.64898","author":[{"given":"Ezekiel","family":"Rozmus","sequence":"first","affiliation":[{"name":"Department of Physiology, University of Kentucky, Lexington, KY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabel G","family":"Stumpf","sequence":"additional","affiliation":[{"name":"Department of Physiology, University of Kentucky, Lexington, KY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Schneider","sequence":"additional","affiliation":[{"name":"Department of Physiology, University of Kentucky, Lexington, KY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander","family":"Alimov","sequence":"additional","affiliation":[{"name":"Department of Physiology, University of Kentucky, Lexington, KY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taylor H.","family":"Pierce","sequence":"additional","affiliation":[{"name":"Department of Physiology, University of Kentucky, Lexington, KY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tooraj","family":"Mirshahi","sequence":"additional","affiliation":[{"name":"Geisinger College of Health Sciences, Danville, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2146-327X","authenticated-orcid":false,"given":"Brian P","family":"Delisle","sequence":"additional","affiliation":[{"name":"Department of Physiology, University of Kentucky, Lexington, KY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7423-5721","authenticated-orcid":false,"given":"Elizabeth A","family":"Schroder","sequence":"additional","affiliation":[{"name":"Department of Physiology, University of Kentucky, Lexington, KY, USA"},{"name":"Department of Internal Medicine, University of Kentucky, Lexington, KY, 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