RRC ID 47486
Author Katoh Y, Terada M, Nishijima Y, Takei R, Nozaki S, Hamada H, Nakayama K.
Title Overall Architecture of the Intraflagellar Transport (IFT)-B Complex Containing Cluap1/IFT38 as an Essential Component of the IFT-B Peripheral Subcomplex.
Journal J Biol Chem
Abstract Intraflagellar transport (IFT) is essential for assembly and maintenance of cilia and flagella as well as ciliary motility and signaling. IFT is mediated by multisubunit complexes, including IFT-A, IFT-B, and the BBSome, in concert with kinesin and dynein motors. Under high salt conditions, purified IFT-B complex dissociates into a core subcomplex composed of at least nine subunits and at least five peripherally associated proteins. Using the visible immunoprecipitation assay, which we recently developed as a convenient protein-protein interaction assay, we determined the overall architecture of the IFT-B complex, which can be divided into core and peripheral subcomplexes composed of 10 and 6 subunits, respectively. In particular, we identified TTC26/IFT56 and Cluap1/IFT38, neither of which was included with certainty in previous models of the IFT-B complex, as integral components of the core and peripheral subcomplexes, respectively. Consistent with this, a ciliogenesis defect of Cluap1-deficient mouse embryonic fibroblasts was rescued by exogenous expression of wild-type Cluap1 but not by mutant Cluap1 lacking the binding ability to other IFT-B components. The detailed interaction map as well as comparison of subcellular localization of IFT-B components between wild-type and Cluap1-deficient cells provides insights into the functional relevance of the architecture of the IFT-B complex.
Volume 291(21)
Pages 10962-75
Published 2016-5-20
DOI 10.1074/jbc.M116.713883
PII S0021-9258(20)40842-7
PMID 26980730
PMC PMC4900248
MeSH Animals Biological Transport Cells, Cultured Flagella / metabolism* HEK293 Cells Humans Immunoprecipitation Intracellular Signaling Peptides and Proteins / chemistry Intracellular Signaling Peptides and Proteins / genetics Intracellular Signaling Peptides and Proteins / metabolism* Kinesins / metabolism Mice Mice, Knockout Multiprotein Complexes / chemistry Multiprotein Complexes / genetics Multiprotein Complexes / metabolism Protein Interaction Maps Protein Subunits
IF 4.238
Times Cited 42
DNA material pCAG-EGFP-C-IFT70B (RDB20234) pEGFP-C1-IFT54 (RDB20235) Genome Network Project Human cDNA Clone IRAK098M05 (HGX039493) IRAK068J01 (HGX027417).