Tissue-specific isoforms of the single C. elegans Ryanodine receptor gene unc-68 control specific functions
Autoři:
Filipe Marques aff001; Saurabh Thapliyal aff001; Avelino Javer aff002; Priyanka Shrestha aff002; André E. X. Brown aff002; Dominique A. Glauser aff001
Působiště autorů:
Department of Biology, University of Fribourg, Fribourg, Switzerland
aff001; MRC London Institute of Medical Sciences, London, United Kingdom
aff002; Institute of Clinical Sciences, Imperial College London, London, United Kingdom
aff003
Vyšlo v časopise:
Tissue-specific isoforms of the single C. elegans Ryanodine receptor gene unc-68 control specific functions. PLoS Genet 16(10): e1009102. doi:10.1371/journal.pgen.1009102
Kategorie:
Research Article
doi:
https://doi.org/10.1371/journal.pgen.1009102
Souhrn
Ryanodine receptors (RyR) are essential regulators of cellular calcium homeostasis and signaling. Vertebrate genomes contain multiple RyR gene isoforms, expressed in different tissues and executing different functions. In contrast, invertebrate genomes contain a single RyR-encoding gene and it has long been proposed that different transcripts generated by alternative splicing may diversify their functions. Here, we analyze the expression and function of alternative exons in the C. elegans RyR gene unc-68. We show that specific isoform subsets are created via alternative promoters and via alternative splicing in unc-68 Divergent Region 2 (DR2), which actually corresponds to a region of high sequence variability across vertebrate isoforms. The expression of specific unc-68 alternative exons is enriched in different tissues, such as in body wall muscle, neurons and pharyngeal muscle. In order to infer the function of specific alternative promoters and alternative exons of unc-68, we selectively deleted them by CRISPR/Cas9 genome editing. We evaluated pharyngeal function, as well as locomotor function in swimming and crawling with high-content computer-assisted postural and behavioral analysis. Our data provide a comprehensive map of the pleiotropic impact of isoform-specific mutations and highlight that tissue-specific unc-68 isoforms fulfill distinct functions. As a whole, our work clarifies how the C. elegans single RyR gene unc-68 can fulfill multiple tasks through tissue-specific isoforms, and provide a solid foundation to further develop C. elegans as a model to study RyR channel functions and malfunctions.
Klíčová slova:
Alternative splicing – Caenorhabditis elegans – Calcium signaling – Invertebrate genomics – Neurons – Pharyngeal muscles – Phenotypes – Swimming
Zdroje
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