Abstract
The human TESTIN (TES) is a putative tumor suppressor and localizes to the cytoplasm as a component of focal adhesions and cell contacts. TES contains a PET domain in the NH2-terminus and three tandem LIM domains in the COOH-terminus. It has been hypothesized that interactions between two termini of TES might lead to a “closed” conformational state of the protein. Here, we provide evidence for different conformational states of TES. We confirmed that the NH2-terminus of TES can interact with its third LIM domain in the COOH-terminus by GST pull-down assays. In addition, antisera against the full-length or two truncations of TES were prepared to examine the relationship between the conformation and cellular distribution of the protein. We found that these antisera recognize different regions of TES and showed that TES is co-localised with the marker protein B23 in nucleolus, in addition to its localization in endoplasmic reticulum (ER). Furthermore, our co-immunoprecipitation (co-IP) analysis of TES and B23 demonstrated their co-existence in the same complex. Taken together, our results suggest that TES has different conformational states in different cellular compartments, and a “closed” conformational state of TES may be involved in nucleolar localization.
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Acknowledgements
This work was supported in part by the National Natural Science Foundation of China (Nos. 20335020, 30470945, 30571005), the Program for Changjiang Scholars and Innovative Research Team in University (No. IRT0445), the 973 project of Ministry of Science and Technology of China (No. 2005CB522505 and 2006CB0F0306), and the Cultivation Fund of the Key Scientific and Technical Innovation Project, Ministry of Education of China (No. 705041).
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Yingli Zhong, Jiaolian Zhu, and Yan Wang contributed equally to the work.
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Zhong, Y., Zhu, J., Wang, Y. et al. LIM domain protein TES changes its conformational states in different cellular compartments. Mol Cell Biochem 320, 85–92 (2009). https://doi.org/10.1007/s11010-008-9901-7
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DOI: https://doi.org/10.1007/s11010-008-9901-7