The Ras suppressor Rsu-1 binds to the LIM 5 domain of the adaptor protein PINCH1 and participates in adhesion-related functions

doi:https://doi.org/10.1016/j.yexcr.2005.01.025

Title: The Ras suppressor Rsu-1 binds to the LIM 5 domain of the adaptor protein PINCH1 and participates in adhesion-related functions

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Abstract

Rsu-1 is a highly conserved leucine rich repeat (LRR) protein that is expressed ubiquitously in mammalian cells. Rsu-1 was identified based on its ability to inhibit transformation by Ras, and previous studies demonstrated that ectopic expression of Rsu-1 inhibited anchorage-independent growth of Ras-transformed cells and human tumor cell lines. Using GAL4-based yeast two-hybrid screening, the LIM domain protein, PINCH1, was identified as the binding partner of Rsu-1. PINCH1 is an adaptor protein that localizes to focal adhesions and it has been implicated in the regulation of adhesion functions. Subdomain mapping in yeast revealed that Rsu-1 binds to the LIM 5 domain of PINCH1, a region not previously identified as a specific binding domain for any other protein. Additional testing demonstrated that PINCH2, which is highly homologous to PINCH1, except in the LIM 5 domain, does not interact with Rsu-1. Glutathione transferase fusion protein binding studies determined that the LRR region of Rsu-1 interacts with PINCH1. Transient expression studies using epitope-tagged Rsu-1 and PINCH1 revealed that Rsu-1 co-immunoprecipitated with PINCH1 and colocalized with vinculin at sites of focal adhesions in mammalian cells. In addition, endogenous P33 Rsu-1 from 293T cells co-immunoprecipitated with transiently expressed myc-tagged PINCH1. Furthermore, RNAi-induced reduction in Rsu-1more »« less

Authors:

Dougherty, Gerard W ^[1]; Chopp, Treasa ^[1]; Shengmei, Qi ^[1]; Cutler, Mary Lou ^[1]

Department of Pathology, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, B3122, Bethesda, MD 20814 (United States)

Publication Date:: 2005-05-15

OSTI Identifier:: 20717603

Resource Type:: Journal Article

Journal Name:: Experimental Cell Research

Additional Journal Information:: Journal Volume: 306; Journal Issue: 1; Other Information: DOI: 10.1016/j.yexcr.2005.01.025; PII: S0014-4827(05)00041-8; Copyright (c) 2005 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0014-4827

Country of Publication:: United States

Language:: English

Subject:: 60 APPLIED LIFE SCIENCES; BIOLOGICAL STRESS; GENE REGULATION; GLUTATHIONE; GROWTH; HYBRIDIZATION; LEUCINE; PHOSPHOTRANSFERASES; RNA; TUMOR CELLS; YEASTS

Citation Formats


                    Dougherty, Gerard W, Section on Structural Cell Biology, National Institute on Deafness and Communication Disorders, Chopp, Treasa, Shengmei, Qi, and Cutler, Mary Lou. The Ras suppressor Rsu-1 binds to the LIM 5 domain of the adaptor protein PINCH1 and participates in adhesion-related functions.  United States: N. p., 2005. 
        Web.  doi:10.1016/j.yexcr.2005.01.025.

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                    Dougherty, Gerard W, Section on Structural Cell Biology, National Institute on Deafness and Communication Disorders, Chopp, Treasa, Shengmei, Qi, & Cutler, Mary Lou. The Ras suppressor Rsu-1 binds to the LIM 5 domain of the adaptor protein PINCH1 and participates in adhesion-related functions.  United States.  https://doi.org/10.1016/j.yexcr.2005.01.025

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                    Dougherty, Gerard W, Section on Structural Cell Biology, National Institute on Deafness and Communication Disorders, Chopp, Treasa, Shengmei, Qi, and Cutler, Mary Lou. Sun .  
        "The Ras suppressor Rsu-1 binds to the LIM 5 domain of the adaptor protein PINCH1 and participates in adhesion-related functions".  United States.  https://doi.org/10.1016/j.yexcr.2005.01.025.

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@article{osti_20717603,

  title        = {The Ras suppressor Rsu-1 binds to the LIM 5 domain of the adaptor protein PINCH1 and participates in adhesion-related functions},

  author       = {Dougherty, Gerard W and Section on Structural Cell Biology, National Institute on Deafness and Communication Disorders and Chopp, Treasa and Shengmei, Qi and Cutler, Mary Lou},

  abstractNote = {Rsu-1 is a highly conserved leucine rich repeat (LRR) protein that is expressed ubiquitously in mammalian cells. Rsu-1 was identified based on its ability to inhibit transformation by Ras, and previous studies demonstrated that ectopic expression of Rsu-1 inhibited anchorage-independent growth of Ras-transformed cells and human tumor cell lines. Using GAL4-based yeast two-hybrid screening, the LIM domain protein, PINCH1, was identified as the binding partner of Rsu-1. PINCH1 is an adaptor protein that localizes to focal adhesions and it has been implicated in the regulation of adhesion functions. Subdomain mapping in yeast revealed that Rsu-1 binds to the LIM 5 domain of PINCH1, a region not previously identified as a specific binding domain for any other protein. Additional testing demonstrated that PINCH2, which is highly homologous to PINCH1, except in the LIM 5 domain, does not interact with Rsu-1. Glutathione transferase fusion protein binding studies determined that the LRR region of Rsu-1 interacts with PINCH1. Transient expression studies using epitope-tagged Rsu-1 and PINCH1 revealed that Rsu-1 co-immunoprecipitated with PINCH1 and colocalized with vinculin at sites of focal adhesions in mammalian cells. In addition, endogenous P33 Rsu-1 from 293T cells co-immunoprecipitated with transiently expressed myc-tagged PINCH1. Furthermore, RNAi-induced reduction in Rsu-1 RNA and protein inhibited cell attachment, and while previous studies demonstrated that ectopic expression of Rsu-1 inhibited Jun kinase activation, the depletion of Rsu-1 resulted in activation of Jun and p38 stress kinases. These studies demonstrate that Rsu-1 interacts with PINCH1 in mammalian cells and functions, in part, by altering cell adhesion.},

  doi          = {10.1016/j.yexcr.2005.01.025},

  url          = {https://www.osti.gov/biblio/20717603},
  journal      = {Experimental Cell Research},

  issn         = {0014-4827},

  number       = 1,

  volume       = 306,

  place        = {United States},

  year         = {2005},

  month        = {5}

}

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