RRC ID 45941
著者 Cevik S, Sanders AA, Van Wijk E, Boldt K, Clarke L, van Reeuwijk J, Hori Y, Horn N, Hetterschijt L, Wdowicz A, Mullins A, Kida K, Kaplan OI, van Beersum SE, Man Wu K, Letteboer SJ, Mans DA, Katada T, Kontani K, Ueffing M, Roepman R, Kremer H, Blacque OE.
タイトル Active transport and diffusion barriers restrict Joubert Syndrome-associated ARL13B/ARL-13 to an Inv-like ciliary membrane subdomain.
ジャーナル PLoS Genet
Abstract Cilia are microtubule-based cell appendages, serving motility, chemo-/mechano-/photo- sensation, and developmental signaling functions. Cilia are comprised of distinct structural and functional subregions including the basal body, transition zone (TZ) and inversin (Inv) compartments, and defects in this organelle are associated with an expanding spectrum of inherited disorders including Bardet-Biedl syndrome (BBS), Meckel-Gruber Syndrome (MKS), Joubert Syndrome (JS) and Nephronophthisis (NPHP). Despite major advances in understanding ciliary trafficking pathways such as intraflagellar transport (IFT), how proteins are transported to subciliary membranes remains poorly understood. Using Caenorhabditis elegans and mammalian cells, we investigated the transport mechanisms underlying compartmentalization of JS-associated ARL13B/ARL-13, which we previously found is restricted at proximal ciliary membranes. We now show evolutionary conservation of ARL13B/ARL-13 localisation to an Inv-like subciliary membrane compartment, excluding the TZ, in many C. elegans ciliated neurons and in a subset of mammalian ciliary subtypes. Compartmentalisation of C. elegans ARL-13 requires a C-terminal RVVP motif and membrane anchoring to prevent distal cilium and nuclear targeting, respectively. Quantitative imaging in more than 20 mutants revealed differential contributions for IFT and ciliopathy modules in defining the ARL-13 compartment; IFT-A/B, IFT-dynein and BBS genes prevent ARL-13 accumulation at periciliary membranes, whereas MKS/NPHP modules additionally inhibit ARL-13 association with TZ membranes. Furthermore, in vivo FRAP analyses revealed distinct roles for IFT and MKS/NPHP genes in regulating a TZ barrier to ARL-13 diffusion, and intraciliary ARL-13 diffusion. Finally, C. elegans ARL-13 undergoes IFT-like motility and quantitative protein complex analysis of human ARL13B identified functional associations with IFT-B complexes, mapped to IFT46 and IFT74 interactions. Together, these findings reveal distinct requirements for sequence motifs, IFT and ciliopathy modules in defining an ARL-13 subciliary membrane compartment. We conclude that MKS/NPHP modules comprise a TZ barrier to ARL-13 diffusion, whereas IFT genes predominantly facilitate ARL-13 ciliary entry and/or retention via active transport mechanisms.
巻・号 9(12)
ページ e1003977
公開日 2013-1-1
DOI 10.1371/journal.pgen.1003977
PII PGENETICS-D-13-00134
PMID 24339792
PMC PMC3854969
MeSH ADP-Ribosylation Factors / genetics* ADP-Ribosylation Factors / metabolism Abnormalities, Multiple Animals Bardet-Biedl Syndrome / genetics Bardet-Biedl Syndrome / pathology Biological Transport, Active / genetics Caenorhabditis elegans / genetics* Caenorhabditis elegans / metabolism Cerebellar Diseases / genetics* Cerebellar Diseases / pathology Cerebellum / abnormalities Cilia / genetics* Cilia / metabolism Ciliary Motility Disorders / genetics Ciliary Motility Disorders / pathology Cytoskeletal Proteins / genetics Cytoskeletal Proteins / metabolism Encephalocele / genetics Encephalocele / pathology Eye Abnormalities / genetics* Eye Abnormalities / pathology Humans Kidney Diseases, Cystic / genetics* Kidney Diseases, Cystic / pathology Membranes / metabolism Polycystic Kidney Diseases / genetics Polycystic Kidney Diseases / pathology Retina / abnormalities* Retina / pathology Retinitis Pigmentosa Transcription Factors / genetics Transcription Factors / metabolism
IF 5.175
引用数 57
WOS 分野 GENETICS & HEREDITY
リソース情報
線虫 tm2322 tm324 tm3100 tm2452 tm925