TY - JOUR
T1 - WDR35 is involved in subcellular localization of acetylated tubulin in 293T cells
AU - Sekiguchi, Takeshi
AU - Ishii, Takashi
AU - Kobayashi, Hideki
AU - Furuno, Nobuaki
N1 - Funding Information:
We appreciate the technical assistance from The Research Support Center, Research Center for Human Disease Modeling, Kyushu University Graduate School of Medical Sciences. This work was supported by JSPS KAKENHI Grant Numbers JP 21570007 ; JP 20K06495 .
Funding Information:
In a previous study, we found that RagA interacts with WDR35 [15]. This finding suggests that, in addition to its function in lysosomes, RagA is involved in the formation or function of primary cilia and in the mTOR signaling process via its interaction with WDR35. In this study, we found that WDR35 interacts with primary cilium-related proteins including the CCT complex, suggesting that WDR35 may play a role in primary cilium formation. WDR35 harbors WD40 repeats and may act as a scaffold protein. As the CCT complex is involved in the formation of primary cilia via assembly of the BBSome [17], the CCT complex may be recruited to primary cilia by WDR35 or may require WDR35 to support its activity in primary cilia. Alternatively, the CCT complex may cause WDR35 to adopt an appropriate higher-order structure for assembly into the IFT-A complex.We appreciate the technical assistance from The Research Support Center, Research Center for Human Disease Modeling, Kyushu University Graduate School of Medical Sciences. This work was supported by JSPS KAKENHI Grant Numbers JP21570007; JP20K06495.
Publisher Copyright:
© 2021 Elsevier Inc.
PY - 2021/4/2
Y1 - 2021/4/2
N2 - WDR35/IFT121 is an intraflagellar transport protein in primary cilia, which is associated with RagA, an mTORC1-activating protein. To elucidate the functions of the interaction between WDR35 and RagA in primary cilia, as well as mTOR signaling, we identified WDR35-interacting proteins using mass spectrometry. We found that WDR35 associates with CCT complex proteins including TCP1/CCT1, which act as molecular chaperones for α-tubulin folding. Immunostaining showed that acetylated α-tubulin was concentrated in the vicinity of primary cilia in 293T cells. In contrast, acetylated tubulin was dispersed in WDR35 partial knockout cells established from 293T cells. Similarly, scattered subcellular localization of acetylated tubulin was observed in RagA knockout cells. RagA was present in the primary cilia of NIH3T3 cells, and the GDP form of RagA exhibited strong binding to WDR35 and negative regulation of primary cilium formation. These results suggest that WDR35 is involved in the subcellular localization of acetylated tubulin in primary cilia via its interactions with TCP1 and/or RagA family proteins.
AB - WDR35/IFT121 is an intraflagellar transport protein in primary cilia, which is associated with RagA, an mTORC1-activating protein. To elucidate the functions of the interaction between WDR35 and RagA in primary cilia, as well as mTOR signaling, we identified WDR35-interacting proteins using mass spectrometry. We found that WDR35 associates with CCT complex proteins including TCP1/CCT1, which act as molecular chaperones for α-tubulin folding. Immunostaining showed that acetylated α-tubulin was concentrated in the vicinity of primary cilia in 293T cells. In contrast, acetylated tubulin was dispersed in WDR35 partial knockout cells established from 293T cells. Similarly, scattered subcellular localization of acetylated tubulin was observed in RagA knockout cells. RagA was present in the primary cilia of NIH3T3 cells, and the GDP form of RagA exhibited strong binding to WDR35 and negative regulation of primary cilium formation. These results suggest that WDR35 is involved in the subcellular localization of acetylated tubulin in primary cilia via its interactions with TCP1 and/or RagA family proteins.
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U2 - 10.1016/j.bbrc.2021.01.092
DO - 10.1016/j.bbrc.2021.01.092
M3 - Article
C2 - 33610917
AN - SCOPUS:85100874268
SN - 0006-291X
VL - 547
SP - 169
EP - 175
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
ER -