title: The conserved SNARE SEC-22 localizes to late endosomes and negatively regulates RNA interference inCaenorhabditis elegans.
abstract: Small RNA pathways, including RNA interference (RNAi), play crucial roles in regulation of gene expression. Initially considered to be cytoplasmic, these processes have later been demonstrated to associate with membranes. For example, maturation of late endosomes/multivesicular bodies (MVBs) is required for efficient RNAi, whereas fusion of MVBs to lysosomes appears to reduce silencing efficiency. SNAREs (solubleN-ethylmaleimide-sensitive factor attachment protein receptors) mediate membrane fusion and are thus at the core of membrane trafficking. In spite of this, no SNARE has previously been reported to affect RNAi. Here, we demonstrate that inCaenorhabditis elegans, loss of the conserved SNARE SEC-22 results in enhanced RNAi upon ingestion of double-stranded RNA. Furthermore, SEC-22 overexpression inhibits RNAi in wild-type animals. We find that overexpression of SEC-22 in the target tissue (body wall muscle) strongly suppresses thesec-22(-)enhanced RNAi phenotype, supporting a primary role for SEC-22 in import of RNAi silencing signals or cell autonomous RNAi. A functional mCherry::SEC-22 protein localizes primarily to late endosomes/MVBs and these compartments are enlarged in animals lackingsec-22SEC-22 interacts with late endosome-associated RNA transport protein SID-5 in a yeast two-hybrid assay and functions in asid-5-dependent manner. Taken together, our data indicate that SEC-22 reduces RNAi efficiency by affecting late endosome/MVB function, for example, by promoting fusion between late endosomes/MVBs and lysosomes. To our knowledge, this is the first report of a SNARE with a function in small RNA-mediated gene silencing.
authors.affiliation: Department of Cell and Molecular Biology, Uppsala University, 751 24 Uppsala, Sweden.
authors.forename: Yani
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Department of Cell and Molecular Biology, Uppsala University, 751 24 Uppsala, Sweden.
authors.forename: Benjamin T
authors.initials: BT
authors.lastname: Holmgren
authors.affiliation: Department of Cell and Molecular Biology, Uppsala University, 751 24 Uppsala, Sweden.
authors.forename: Andrea
authors.identifier: 0000-0002-9353-0742
authors.initials: A
authors.lastname: Hinas
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics* / Q000378:metabolism; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism*; D017930:Genes, Reporter; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism; D008561:Membrane Fusion; D011505:Protein-Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics* / Q000378:metabolism; D018510:RNA, Helminth / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D050600:SNARE Proteins / Q000235:genetics* / Q000378:metabolism; D000097573:Red Fluorescent Protein
chemical_list: D029742:Caenorhabditis elegans Proteins; D008164:Luminescent Proteins; D011505:Protein-Tyrosine Kinases; D012330:RNA, Double-Stranded; D018510:RNA, Helminth; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D050600:SNARE Proteins; D000097573:Red Fluorescent Protein; C000622350:SEC-22 protein, C elegans; C000622353:sid-5 protein, C elegans
keywords: C. elegans; RNAi; SNARE; late endosome