p104 Binds to Rac1 and Reduces Its Activity during Myotube Differentiation of C2C12 Cell
The p104 protein inhibits cellular proliferation when overexpressed in NIH3T3 cells and has been shown to associate with p85α, Grb2, and PLCγ1. In order to isolate other proteins that interact with p104, yeast two-hybrid screening was performed. Rac1 was identified as a binding partner of p104 and t...
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| Format: | Article |
| Language: | English |
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Wiley
2014-01-01
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| Series: | The Scientific World Journal |
| Online Access: | http://dx.doi.org/10.1155/2014/592450 |
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| _version_ | 1832551635546537984 |
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| author | Ki Young Choi Min Sup Lee Young Jun Cho Myong Ho Jeong Seung Jin Han Seung Hwan Hong |
| author_facet | Ki Young Choi Min Sup Lee Young Jun Cho Myong Ho Jeong Seung Jin Han Seung Hwan Hong |
| author_sort | Ki Young Choi |
| collection | DOAJ |
| description | The p104 protein inhibits cellular proliferation when overexpressed in NIH3T3 cells and has been shown to associate with p85α, Grb2, and PLCγ1. In order to isolate other proteins that interact with p104, yeast two-hybrid screening was performed. Rac1 was identified as a binding partner of p104 and the interaction between p104 and Rac1 was confirmed by immunoprecipitation. Using a glutathione S-transferase (GST) pull-down assay with various p104 fragments, the 814–848 amino acid residue at the carboxyl-terminal region of p104 was identified as the key component to interact with Rac1. The CrkII which is involved in the Rac1-mediated cellular response was also found to interact with p104 protein. NIH3T3 cells which overexpressed p104 showed a decrease of Rac1 activity. However, neither the proline-rich domain mutant, which is unable to interact with CrkII, nor the carboxy-terminal deletion mutant could attenuate Rac1 activity. During the differentiation of myoblasts, the amount of p104 protein as well as transcript level was increased. The overexpression of p104 enhanced myotube differentiation, whereas siRNA of p104 reversed this process. In this process, more Rac1 and CrkII were bound to increased p104. Based on these results, we conclude that p104 is involved in muscle cell differentiation by modulating the Rac1 activity. |
| format | Article |
| id | doaj-art-d5cae01486844b199c9f4fe53dfa533d |
| institution | Kabale University |
| issn | 2356-6140 1537-744X |
| language | English |
| publishDate | 2014-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | The Scientific World Journal |
| spelling | doaj-art-d5cae01486844b199c9f4fe53dfa533d2025-02-03T06:01:08ZengWileyThe Scientific World Journal2356-61401537-744X2014-01-01201410.1155/2014/592450592450p104 Binds to Rac1 and Reduces Its Activity during Myotube Differentiation of C2C12 CellKi Young Choi0Min Sup Lee1Young Jun Cho2Myong Ho Jeong3Seung Jin Han4Seung Hwan Hong5Institute of Molecular Biology and Genetics, Seoul National University, Seoul 151-742, Republic of KoreaInstitute of Molecular Biology and Genetics, Seoul National University, Seoul 151-742, Republic of KoreaSchool of Biological Sciences, Seoul National University, San 56-1 Shillim-dong, Kwanak-gu, Seoul 151-742, Republic of KoreaSchool of Biological Sciences, Inje University, Gimhae 621-749, Republic of KoreaSchool of Biological Sciences, Inje University, Gimhae 621-749, Republic of KoreaInstitute of Molecular Biology and Genetics, Seoul National University, Seoul 151-742, Republic of KoreaThe p104 protein inhibits cellular proliferation when overexpressed in NIH3T3 cells and has been shown to associate with p85α, Grb2, and PLCγ1. In order to isolate other proteins that interact with p104, yeast two-hybrid screening was performed. Rac1 was identified as a binding partner of p104 and the interaction between p104 and Rac1 was confirmed by immunoprecipitation. Using a glutathione S-transferase (GST) pull-down assay with various p104 fragments, the 814–848 amino acid residue at the carboxyl-terminal region of p104 was identified as the key component to interact with Rac1. The CrkII which is involved in the Rac1-mediated cellular response was also found to interact with p104 protein. NIH3T3 cells which overexpressed p104 showed a decrease of Rac1 activity. However, neither the proline-rich domain mutant, which is unable to interact with CrkII, nor the carboxy-terminal deletion mutant could attenuate Rac1 activity. During the differentiation of myoblasts, the amount of p104 protein as well as transcript level was increased. The overexpression of p104 enhanced myotube differentiation, whereas siRNA of p104 reversed this process. In this process, more Rac1 and CrkII were bound to increased p104. Based on these results, we conclude that p104 is involved in muscle cell differentiation by modulating the Rac1 activity.http://dx.doi.org/10.1155/2014/592450 |
| spellingShingle | Ki Young Choi Min Sup Lee Young Jun Cho Myong Ho Jeong Seung Jin Han Seung Hwan Hong p104 Binds to Rac1 and Reduces Its Activity during Myotube Differentiation of C2C12 Cell The Scientific World Journal |
| title | p104 Binds to Rac1 and Reduces Its Activity during Myotube Differentiation of C2C12 Cell |
| title_full | p104 Binds to Rac1 and Reduces Its Activity during Myotube Differentiation of C2C12 Cell |
| title_fullStr | p104 Binds to Rac1 and Reduces Its Activity during Myotube Differentiation of C2C12 Cell |
| title_full_unstemmed | p104 Binds to Rac1 and Reduces Its Activity during Myotube Differentiation of C2C12 Cell |
| title_short | p104 Binds to Rac1 and Reduces Its Activity during Myotube Differentiation of C2C12 Cell |
| title_sort | p104 binds to rac1 and reduces its activity during myotube differentiation of c2c12 cell |
| url | http://dx.doi.org/10.1155/2014/592450 |
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