2024-03-29T02:12:56Z
https://nagoya.repo.nii.ac.jp/oai
oai:nagoya.repo.nii.ac.jp:00024073
2023-01-16T04:54:37Z
643:666:667
The calcium-binding protein ALG-2 promotes endoplasmic reticulum exit site localization and polymerization of Trk-fused gene (TFG) protein
Kanadome, Takashi
Shibata, Hideki
Kuwata, Keiko
Takahara, Terunao
Maki, Masatoshi
open access
This is the peer reviewed version of the following article: [Kanadome, T., Shibata, H., Kuwata, K., Takahara, T. and Maki, M. (2017), The calcium-binding protein ALG-2 promotes endoplasmic reticulum exit site localization and polymerization of Trk-fused gene (TFG) protein. FEBS J, 284: 56–76. doi:10.1111/febs.13949], which has been published in final form at [http://doi.org/10.1111/febs.13949]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.
Apoptosis-linked gene 2 (ALG-2), which is a gene product of PDCD6, is a 22-kDa Ca2+-binding protein. Accumulating evidence points to a role for ALG-2 as a Ca2+-responsive adaptor protein. On binding to Ca2+, ALG-2 undergoes a conformational change that facilitates its interaction with various proteins. It also forms a homodimer and heterodimer with peflin, a paralog of ALG-2. However, the differences in cellular roles for the ALG-2 homodimer and ALG-2/peflin heterodimer are unclear. In the present study, we found that Trk-fused gene (TFG) protein interacted with the ALG-2 homodimer. Immunostaining analysis revealed that TFG and ALG-2 partially overlapped at endoplasmic reticulum exit sites (ERES), a platform for COPII-mediated protein transport from the endoplasmic reticulum. Time-lapse live-cell imaging demonstrated that both green fluorescent protein-fused TFG and mCherry-fused ALG-2 are recruited to ERES after thapsigargin treatment, which raises intracellular Ca2+ levels. Furthermore, overexpression of ALG-2 induced the accumulation of TFG at ERES. TFG has an ALG-2-binding motif and deletion of the motif decreased TFG binding to ALG-2 and shortened its half-life at ERES, suggesting a critical role for ALG-2 in retaining TFG at ERES. We also demonstrated, by in vitro cross-linking assays, that ALG-2 promoted the polymerization of TFG in a Ca2+-dependent manner. Collectively, the results suggest that ALG-2 acts as a Ca2+-sensitive adaptor to concentrate and polymerize TFG at ERES, supporting a potential role for ALG-2 in COPII-dependent trafficking from the endoplasmic reticulum.
Wiley
2017-01
eng
journal article
AM
http://hdl.handle.net/2237/26285
https://nagoya.repo.nii.ac.jp/records/24073
https://doi.org/10.1111/febs.13949
1742-464X
FEBS Journal
284
1
56
76
https://nagoya.repo.nii.ac.jp/record/24073/files/FJ_KanadomeT_ShibataH_170510.pdf
application/pdf
7.5 MB
2018-01-01