{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T02:43:23Z","timestamp":1768531403621,"version":"3.49.0"},"posted":{"date-parts":[[2025,11,16]]},"group-title":"Developmental Biology","reference-count":101,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,11,16]],"date-time":"2025-11-16T00:00:00Z","timestamp":1763251200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,11,16]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Apical extracellular matrices (aECMs) are associated with many epithelia and many form a protective layer against biotic and abiotic threats in the environment. Despite their importance, we lack a deep understanding of their structure and dynamics in development and disease.\n                  <jats:italic>C. elegans<\/jats:italic>\n                  molting offers a powerful entry point to understanding developmentally programmed aECM remodeling. Here, we show that the poorly characterized putative protease inhibitor gene,\n                  <jats:italic>mlt-11<\/jats:italic>\n                  , is directly regulated by the NHR-23 transcription factor. We identify key\n                  <jats:italic>cis-<\/jats:italic>\n                  regulatory elements required for robust\n                  <jats:italic>mlt-11<\/jats:italic>\n                  expression. An internal MLT-11::mNeonGreen translational fusion transiently localized to the aECM in the cuticle and embryo. MLT-11::mNeonGreen was also detected in lining openings to the exterior (vulva, rectum, mouth).\n                  <jats:italic>mlt-11<\/jats:italic>\n                  is necessary to pattern all layers of the adult cuticle, and reduction of MLT-11 levels disrupted the barrier function of the cuticle. Deletion of conserved Kunitz protease inhibitor domains or intervening sequences produced a range of defects including either left or right rollers, and small separations of the cuticle along the length of the animal (microblisters). MLT-11 is processed into at least two fragments and internal and C-terminal mNeonGreen knock-ins display distinct localization patterns. Predicted\n                  <jats:italic>mlt-11<\/jats:italic>\n                  null mutations caused fully penetrant embryonic lethality and elongation defects. Together, this work suggests that MLT-11 localizes similarly to pre-cuticle components and conserved sequences play distinct roles in promoting proper assembly of the aECM.\n                <\/jats:p>","DOI":"10.1101\/2025.11.14.688552","type":"posted-content","created":{"date-parts":[[2025,11,16]],"date-time":"2025-11-16T23:55:14Z","timestamp":1763337314000},"source":"Crossref","is-referenced-by-count":0,"title":["MLT-11 is necessary for\n                  <i>C. elegans<\/i>\n                  embryogenesis and conserved sequences play distinct roles in cuticle structure"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6626-2615","authenticated-orcid":false,"given":"James Matthew","family":"Ragle","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ariela","family":"Turzo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anton","family":"Jackson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"An A.","family":"Vo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vivian T.","family":"Pham","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Keya","family":"Daly","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2867-9740","authenticated-orcid":false,"given":"John C.","family":"Clancy","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Max T.","family":"Levenson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alex D.","family":"Lee","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9870-8936","authenticated-orcid":false,"given":"Jordan D.","family":"Ward","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025111818051237000_2025.11.14.688552v1.1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-43058-9"},{"key":"2025111818051237000_2025.11.14.688552v1.2","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.75906"},{"key":"2025111818051237000_2025.11.14.688552v1.3","doi-asserted-by":"publisher","DOI":"10.2174\/1389203033487180"},{"key":"2025111818051237000_2025.11.14.688552v1.4","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/iyab006"},{"key":"2025111818051237000_2025.11.14.688552v1.5","unstructured":"Bada Juarez J et al. 2019. Lipodisqs for eukaryote lipidomics with retention of viability: Sensitivity and resistance to Leucobacter infection linked to C.elegans cuticle composition. Chem Phys Lipids. 222(2019) [accessed 2024 May 9]. 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