RRC ID 53819
Author Hamada Y, Tsurumi Y, Nozaki S, Katoh Y, Nakayama K.
Title Interaction of WDR60 intermediate chain with TCTEX1D2 light chain of the dynein-2 complex is crucial for ciliary protein trafficking.
Journal Mol Biol Cell
Abstract The dynein-2 complex mediates trafficking of ciliary proteins by powering the intraflagellar transport (IFT) machinery containing IFT-A and IFT-B complexes. Although 11 subunits are known to constitute the dynein-2 complex, with several light-chain subunits shared by the dynein-1 complex, the overall architecture of the dynein-2 complex has not been fully clarified. Utilizing the visible immunoprecipitation assay, we demonstrated the interaction modes among the dynein-2 subunits, including previously undefined interactions, such as that between WDR60 and the TCTEX1D2-DYNLT1/DYNLT3 dimer. The dynein-2 complex can be divided into three subcomplexes, namely DYNC2H1-DYNC2LI1, WDR34-DYNLL1/DYNLL2-DYNLRB1/DYNLRB2, and WDR60-TCTEX1D2-DYNLT1/DYNLT3. We established cell lines lacking WDR60 or TCTEX1D2, both of which are dynein-2-specific subunits encoded by ciliopathy-causing genes, and found that both WDR60-knockout (KO) and TCTEX1D2-KO cells show defects in retrograde ciliary protein trafficking, with WDR60-KO cells demonstrating more severe defects probably due to failed assembly of the dynein-2 complex. The exogenous expression of a WDR60 mutant lacking TCTEX1D2 binding partially restored retrograde trafficking to a level comparable to that of TCTEX1D2-KO cells. Thus, our results demonstrated that WDR60 plays a major role and TCTEX1D2 plays an auxiliary role in the dynein-2 complex to mediate retrograde ciliary protein trafficking.
Volume 29(13)
Pages 1628-1639
Published 2018-7-1
DOI 10.1091/mbc.E18-03-0173
PMID 29742051
PMC PMC6080652
MeSH Adaptor Proteins, Signal Transducing / chemistry Adaptor Proteins, Signal Transducing / metabolism* Amino Acid Sequence Biological Assay Cell Line Cilia / metabolism* Conserved Sequence Cytoskeletal Proteins Dyneins / metabolism* Gene Knockdown Techniques HEK293 Cells Humans Protein Binding Protein Domains Protein Multimerization Protein Subunits / metabolism Protein Transport
IF 3.791
Times Cited 9
DNA material pCAG2-EGFP-C-DYNC2LI1 (RDB20239) pCAG2-EGFP-C-WDR34 (RDB20240) pCAG2-EGFP-C-WDR60 (RDB20241) Genome Network Project Human cDNA Clone IRAL016M21 (HGY086709) IRAL028A16 (HGY091216) IRAL034P23 (HGY093983)