{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T12:13:33Z","timestamp":1788956013631,"version":"build-2803163510"},"posted":{"date-parts":[[2026,5,15]]},"group-title":"Developmental Biology","reference-count":121,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,15]],"date-time":"2026-05-15T00:00:00Z","timestamp":1778803200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000001","name":"U.S. National Science Foundation","doi-asserted-by":"crossref","award":["2140001"],"award-info":[{"award-number":["2140001"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"crossref"}]},{"award":["R01 NS118078"],"award-info":[{"award-number":["R01 NS118078"]}],"id":[{"id":"https:\/\/ror.org\/01s5ya894","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01 NS116365"],"award-info":[{"award-number":["R01 NS116365"]}],"id":[{"id":"https:\/\/ror.org\/01s5ya894","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  How post-mitotic neurons maintain precise transcription factor (TF) levels throughout life remains a fundamental open question. Here, we challenge the prevailing model of positive autoregulation by demonstrating that UNC-3 (Collier\/EBF1-4), a dosage-sensitive TF continuously required for cholinergic motor neuron identity in\n                  <jats:italic>C. elegans<\/jats:italic>\n                  , negatively regulates its own expression. Using genetics, biochemistry, and inducible protein depletion, we show this self-repression occurs directly at the transcriptional level and persists beyond development. CRISPR\/Cas9 disruption of negative autoregulation causes motor neuron identity and locomotion defects, establishing its functional necessity. Mechanistically, the UNC-3 DNA-binding domain is required and sufficient for self-repression, with an AlphaFold2 screen implicating chromatin factors as interaction partners. Critically, UNC-3 self-repression is continuously counterbalanced by positive input from the HOX cofactor CEH-20\/PBX, revealing a dynamic \u201cbalancing act\u201d between opposing regulatory inputs that stabilize TF dosage over time. Mutations in the\n                  <jats:italic>unc-3<\/jats:italic>\n                  ortholog\n                  <jats:italic>EBF3<\/jats:italic>\n                  cause a neurodevelopmental syndrome, and disease-associated variants disrupt UNC-3 self-repression, revealing a key molecular mechanism underlying the disorder. We propose that negative autoregulation continuously counteracted by positive input represents a broadly applicable principle for maintaining dosage-sensitive TF expression to secure post-mitotic cell identity.\n                <\/jats:p>","DOI":"10.64898\/2026.05.14.725191","type":"posted-content","created":{"date-parts":[[2026,5,16]],"date-time":"2026-05-16T06:50:13Z","timestamp":1778914213000},"source":"Crossref","is-referenced-by-count":1,"title":["Continuous negative autoregulation fine-tunes dosage-sensitive transcription factor expression to maintain post-mitotic neuron identity"],"prefix":"10.64898","author":[{"given":"Honorine","family":"Destain","sequence":"first","affiliation":[{"name":"Department of Neurobiology, University of Chicago, Chicago, IL, USA"},{"name":"University of Chicago Neuroscience Institute, Chicago, IL, USA"},{"name":"Committee on Development, Regeneration, and Stem Cell Biology, University of Chicago, Chicago, IL, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9267-455X","authenticated-orcid":false,"given":"Alan","family":"Koh","sequence":"additional","affiliation":[{"name":"MRC London Institute of Medical Sciences, London, UK"},{"name":"Institute of Clinical Sciences, Imperial College London, London, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heewhan","family":"Shin","sequence":"additional","affiliation":[{"name":"Department of Biochemistry & Molecular Biology, University of Chicago, Chicago, IL, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e9 E. 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