TY - JOUR TI - A spatiotemporal reconstruction of the C. elegans pharyngeal cuticle reveals a structure rich in phase-separating proteins AU - Kamal, Muntasir AU - Tokmakjian, Levon AU - Knox, Jessica AU - Mastrangelo, Peter AU - Ji, Jingxiu AU - Cai, Hao AU - Wojciechowski, Jakub W AU - Hughes, Michael P AU - Takács, Kristóf AU - Chu, Xiaoquan AU - Pei, Jianfeng AU - Grolmusz, Vince AU - Kotulska, Malgorzata AU - Forman-Kay, Julie Deborah AU - Roy, Peter J A2 - Cochella, Luisa A2 - Sengupta, Piali VL - 11 PY - 2022 DA - 2022/10/19 SP - e79396 C1 - eLife 2022;11:e79396 DO - 10.7554/eLife.79396 UR - https://doi.org/10.7554/eLife.79396 AB - How the cuticles of the roughly 4.5 million species of ecdysozoan animals are constructed is not well understood. Here, we systematically mine gene expression datasets to uncover the spatiotemporal blueprint for how the chitin-based pharyngeal cuticle of the nematode Caenorhabditis elegans is built. We demonstrate that the blueprint correctly predicts expression patterns and functional relevance to cuticle development. We find that as larvae prepare to molt, catabolic enzymes are upregulated and the genes that encode chitin synthase, chitin cross-linkers, and homologs of amyloid regulators subsequently peak in expression. Forty-eight percent of the gene products secreted during the molt are predicted to be intrinsically disordered proteins (IDPs), many of which belong to four distinct families whose transcripts are expressed in overlapping waves. These include the IDPAs, IDPBs, and IDPCs, which are introduced for the first time here. All four families have sequence properties that drive phase separation and we demonstrate phase separation for one exemplar in vitro. This systematic analysis represents the first blueprint for cuticle construction and highlights the massive contribution that phase-separating materials make to the structure. KW - cuticle KW - intrinsically disordered proteins KW - molting KW - systems analyses KW - phase separation KW - novel gene families JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -