Cell–cell adhesion plays crucial roles in cell differentiation and morphogenesis during development of Dictyostelium discoideum. The heterophilic adhesion protein TgrC1 (Tgr is transmembrane, IPT, IG, E-set, repeat protein) is expressed during cell aggregation, and disruption of the tgrC1 gene results in the arrest of development at the loose aggregate stage. We have used far-Western blotting coupled with MS to identify TgrB1 as the heterophilic binding partner of TgrC1. Co-immunoprecipitation and pull-down studies showed that TgrB1 and TgrC1 are capable of binding with each other in solution. TgrB1 and TgrC1 are encoded by a pair of adjacent genes which share a common promoter. Both TgrB1 and TgrC1 are type I transmembrane proteins, which contain three extracellular IPT/TIG (immunoglobulin, plexin, transcription factor-like/transcription factor immunoglobulin) domains. Antibodies raised against TgrB1 inhibit cell reassociation at the post-aggregation stage of development and block fruiting body formation. Ectopic expression of TgrB1 and TgrC1 driven by the actin15 promoter leads to heterotypic cell aggregation of vegetative cells. Using recombinant proteins that cover different portions of TgrB1 and TgrC1 in binding assays, we have mapped the cell-binding regions in these two proteins to Lys537–Ala783 in TgrB1 and Ile336–Val360 in TgrC1, corresponding to their respective TIG3 and TIG2 domain.
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Research Article|
May 10 2013
TgrC1 mediates cell–cell adhesion by interacting with TgrB1 via mutual IPT/TIG domains during development of Dictyostelium discoideum
Gong Chen;
Gong Chen
*Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada, M5S 1A8
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Jun Wang;
Jun Wang
*Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada, M5S 1A8
†School of Biological Science and Technology, Central South University, Changsha, Hunan Province 410078, People's Republic of China
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Xiaoqun Xu;
Xiaoqun Xu
‡Banting and Best Department of Medical Research, University of Toronto, Toronto, Ontario, Canada, M5S 1A8
§Institute of Basic Medicine, Shandong Academy of Medical Sciences, Jinan, Shandong Province 250062, People's Republic of China
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Xiangfu Wu;
Xiangfu Wu
‡Banting and Best Department of Medical Research, University of Toronto, Toronto, Ontario, Canada, M5S 1A8
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Ruihan Piao;
Ruihan Piao
*Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada, M5S 1A8
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Chi-Hung Siu
Chi-Hung Siu
1
*Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada, M5S 1A8
‡Banting and Best Department of Medical Research, University of Toronto, Toronto, Ontario, Canada, M5S 1A8
1To whom correspondence should be addressed (email chi.hung.siu@utoronto.ca).
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Publisher: Portland Press Ltd
Received:
November 01 2012
Revision Received:
March 07 2013
Accepted:
March 12 2013
Accepted Manuscript online:
March 12 2013
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2013 Biochemical Society
2013
Biochem J (2013) 452 (2): 259–269.
Article history
Received:
November 01 2012
Revision Received:
March 07 2013
Accepted:
March 12 2013
Accepted Manuscript online:
March 12 2013
Citation
Gong Chen, Jun Wang, Xiaoqun Xu, Xiangfu Wu, Ruihan Piao, Chi-Hung Siu; TgrC1 mediates cell–cell adhesion by interacting with TgrB1 via mutual IPT/TIG domains during development of Dictyostelium discoideum. Biochem J 1 June 2013; 452 (2): 259–269. doi: https://doi.org/10.1042/BJ20121674
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