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Title: Molecular basis for multimerization in the activation of the epidermal growth factor receptor

Abstract

The epidermal growth factor receptor (EGFR) is activated by dimerization, but activation also generates higher-order multimers, whose nature and function are poorly understood. We have characterized ligand-induced dimerization and multimerization of EGFR using single-molecule analysis, and show that multimerization can be blocked by mutations in a specific region of Domain IV of the extracellular module. These mutations reduce autophosphorylation of the C-terminal tail of EGFR and attenuate phosphorylation of phosphatidyl inositol 3-kinase, which is recruited by EGFR. The catalytic activity of EGFR is switched on through allosteric activation of one kinase domain by another, and we show that if this is restricted to dimers, then sites in the tail that are proximal to the kinase domain are phosphorylated in only one subunit. We propose a structural model for EGFR multimerization through self-association of ligand-bound dimers, in which the majority of kinase domains are activated cooperatively, thereby boosting tail phosphorylation.

Authors:
 [1];  [2];  [3];  [3];  [4];  [4];  [4];  [4]; ORCiD logo [5]; ORCiD logo [6]
  1. Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology, California Inst. for Quantitative Biosciences, Howard Hughes Medical Inst., Biophysics Graduate Group
  2. Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology
  3. Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology, California Inst. for Quantitative Biosciences, Howard Hughes Medical Inst.
  4. Univ. of Akron, OH (United States). Dept. of Chemistry
  5. Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology, California Inst. for Quantitative Biosciences, Biophysics Graduate Group, Dept. of Chemistry, Helen Wills Neuroscience Inst.
  6. Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology, California Inst. for Quantitative Biosciences, Howard Hughes Medical Inst., Biophysics Graduate Group, Dept. of Chemistry; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Physical Biosciences Division
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
National Institutes of Health (NIH); National Cancer Institute (NCI)
OSTI Identifier:
1379248
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
eLife
Additional Journal Information:
Journal Volume: 5; Journal Issue: MARCH2016; Journal ID: ISSN 2050-084X
Publisher:
eLife Sciences Publications, Ltd.
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Huang, Yongjian, Bharill, Shashank, Karandur, Deepti, Peterson, Sean M., Marita, Morgan, Shi, Xiaojun, Kaliszewski, Megan J., Smith, Adam W., Isacoff, Ehud Y., and Kuriyan, John. Molecular basis for multimerization in the activation of the epidermal growth factor receptor. United States: N. p., 2016. Web. doi:10.7554/eLife.14107.
Huang, Yongjian, Bharill, Shashank, Karandur, Deepti, Peterson, Sean M., Marita, Morgan, Shi, Xiaojun, Kaliszewski, Megan J., Smith, Adam W., Isacoff, Ehud Y., & Kuriyan, John. Molecular basis for multimerization in the activation of the epidermal growth factor receptor. United States. doi:10.7554/eLife.14107.
Huang, Yongjian, Bharill, Shashank, Karandur, Deepti, Peterson, Sean M., Marita, Morgan, Shi, Xiaojun, Kaliszewski, Megan J., Smith, Adam W., Isacoff, Ehud Y., and Kuriyan, John. Mon . "Molecular basis for multimerization in the activation of the epidermal growth factor receptor". United States. doi:10.7554/eLife.14107. https://www.osti.gov/servlets/purl/1379248.
@article{osti_1379248,
title = {Molecular basis for multimerization in the activation of the epidermal growth factor receptor},
author = {Huang, Yongjian and Bharill, Shashank and Karandur, Deepti and Peterson, Sean M. and Marita, Morgan and Shi, Xiaojun and Kaliszewski, Megan J. and Smith, Adam W. and Isacoff, Ehud Y. and Kuriyan, John},
abstractNote = {The epidermal growth factor receptor (EGFR) is activated by dimerization, but activation also generates higher-order multimers, whose nature and function are poorly understood. We have characterized ligand-induced dimerization and multimerization of EGFR using single-molecule analysis, and show that multimerization can be blocked by mutations in a specific region of Domain IV of the extracellular module. These mutations reduce autophosphorylation of the C-terminal tail of EGFR and attenuate phosphorylation of phosphatidyl inositol 3-kinase, which is recruited by EGFR. The catalytic activity of EGFR is switched on through allosteric activation of one kinase domain by another, and we show that if this is restricted to dimers, then sites in the tail that are proximal to the kinase domain are phosphorylated in only one subunit. We propose a structural model for EGFR multimerization through self-association of ligand-bound dimers, in which the majority of kinase domains are activated cooperatively, thereby boosting tail phosphorylation.},
doi = {10.7554/eLife.14107},
journal = {eLife},
number = MARCH2016,
volume = 5,
place = {United States},
year = {2016},
month = {3}
}

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