TY - JOUR
T1 - Formation of the B9-domain protein complex MKS1-B9D2-B9D1 is essential as a diffusion barrier for ciliary membrane proteins
AU - Okazaki, Misato
AU - Kobayashi, Takuya
AU - Chiba, Shuhei
AU - Takei, Ryota
AU - Liang, Luxiaoxue
AU - Nakayama, Kazuhisa
AU - Katoh, Yohei
N1 - Funding Information:
We thank Peter McPherson for providing plasmid vectors for recombinant lentivirus production, Shohei Nozaki and Fumiyoshi Ishidate for technical assistance and advice, and Helena Akiko Popiel for critical reading of the manuscript. This work was supported in part by grants from the Japan Society for the Promotion of Science (JSPS) (grant numbers 15H04370 and 19H00980 to K.N. and 15K07929 and 18H02403 to Y.K.), a grant of JRPs-LEAD with UKRI from JSPS, a grant from the Uehara Memorial Foundation to Y.K., and the Kyoto University internal grant ISHIZUE to K.N. We used Airyscan microscopy at the Research Support Platform in Osaka City University and the SH800 Series cell sorter at the Medical Research Support Center in Graduate School of Medicine, Kyoto University.
Publisher Copyright:
© 2020 Okazaki et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution-Noncommercial-Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).
PY - 2020/9
Y1 - 2020/9
N2 - Cilia are plasma membrane protrusions that act as cellular antennae and propellers in eukaryotes. To achieve their sensory and motile functions, cilia maintain protein and lipid compositions that are distinct from those of the cell body. The transition zone (TZ) is a specialized region located at the ciliary base, which functions as a barrier separating the interior and exterior of cilia. The TZ comprises a number of transmembrane and soluble proteins. Meckel syndrome (MKS)1, B9 domain (B9D)1/MKS9, and B9D2/MKS10 are soluble TZ proteins that are encoded by causative genes of MKS and have a B9D in common. We here demonstrate the interaction mode of these B9D proteins to be MKS1-B9D2-B9D1 and demonstrate their interdependent localization to the TZ. Phenotypic analyses of MKS1-knockout (KO) and B9D2-KO cells show that the B9D proteins are involved in, although not essential for, normal cilia biogenesis. Rescue experiments of these KO cells show that formation of the B9D protein complex is crucial for creating a diffusion barrier for ciliary membrane proteins.
AB - Cilia are plasma membrane protrusions that act as cellular antennae and propellers in eukaryotes. To achieve their sensory and motile functions, cilia maintain protein and lipid compositions that are distinct from those of the cell body. The transition zone (TZ) is a specialized region located at the ciliary base, which functions as a barrier separating the interior and exterior of cilia. The TZ comprises a number of transmembrane and soluble proteins. Meckel syndrome (MKS)1, B9 domain (B9D)1/MKS9, and B9D2/MKS10 are soluble TZ proteins that are encoded by causative genes of MKS and have a B9D in common. We here demonstrate the interaction mode of these B9D proteins to be MKS1-B9D2-B9D1 and demonstrate their interdependent localization to the TZ. Phenotypic analyses of MKS1-knockout (KO) and B9D2-KO cells show that the B9D proteins are involved in, although not essential for, normal cilia biogenesis. Rescue experiments of these KO cells show that formation of the B9D protein complex is crucial for creating a diffusion barrier for ciliary membrane proteins.
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U2 - 10.1091/MBC.E20-03-0208
DO - 10.1091/MBC.E20-03-0208
M3 - Article
C2 - 32726168
AN - SCOPUS:85091125761
SN - 1059-1524
VL - 31
SP - 2259
EP - 2268
JO - Molecular Biology of the Cell
JF - Molecular Biology of the Cell
IS - 20
ER -