Regulation of Rho guanine nucleotide exchange factor 3 by phosphorylation in the PH domain
Summary: Although most members of the Rho guanine nucleotide exchange factor (RhoGEF) family are found to be phosphorylated, how phosphorylation regulates RhoGEF function is poorly understood. Here we report the discovery of a mechanism of RhoGEF regulation by phosphorylation. We find that ARHGEF3 i...
Saved in:
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-06-01
|
| Series: | iScience |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004225010144 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849331011896213504 |
|---|---|
| author | Jesus F. Moreno Jae-Sung You Carlos C. Rodriguez Shashank Pant Adriana Reyes-Ordoñez Reean Abdullah Nilmani Singh Maxine J. van der Donk Emad Tajkhorshid Jie Chen |
| author_facet | Jesus F. Moreno Jae-Sung You Carlos C. Rodriguez Shashank Pant Adriana Reyes-Ordoñez Reean Abdullah Nilmani Singh Maxine J. van der Donk Emad Tajkhorshid Jie Chen |
| author_sort | Jesus F. Moreno |
| collection | DOAJ |
| description | Summary: Although most members of the Rho guanine nucleotide exchange factor (RhoGEF) family are found to be phosphorylated, how phosphorylation regulates RhoGEF function is poorly understood. Here we report the discovery of a mechanism of RhoGEF regulation by phosphorylation. We find that ARHGEF3 is phosphorylated in the pleckstrin homology (PH) domain in a protein kinase C (PKC)-dependent manner. This phosphorylation inhibits ARHGEF3 activation of RhoA and actin stress fiber formation in the cells, and it also disrupts ARHGEF3 binding to PI(3,5)P2 but not to PI(4,5)P2. Guided by molecular dynamics simulation, a mutation in the PH domain is identified to uncouple ARHGEF3 binding to the two lipids, which is used to rule out a role of PI(3,5)P2 in regulating GEF activity. Results of in vitro GEF assays suggest that PH domain phosphorylation diminishes ARHGEF3 catalytic activity, likely through an allosteric mechanism. Our findings reveal a previously unknown type of regulatory mechanism for the family of RhoGEFs. |
| format | Article |
| id | doaj-art-42b5e8e43a13436d946dd85ebc7e84dc |
| institution | Kabale University |
| issn | 2589-0042 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj-art-42b5e8e43a13436d946dd85ebc7e84dc2025-08-20T03:46:45ZengElsevieriScience2589-00422025-06-0128611275310.1016/j.isci.2025.112753Regulation of Rho guanine nucleotide exchange factor 3 by phosphorylation in the PH domainJesus F. Moreno0Jae-Sung You1Carlos C. Rodriguez2Shashank Pant3Adriana Reyes-Ordoñez4Reean Abdullah5Nilmani Singh6Maxine J. van der Donk7Emad Tajkhorshid8Jie Chen9Department of Cell and Developmental Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USADepartment of Cell and Developmental Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USADepartment of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Theoretical and Computational Biophysics Group, NIH Resource for Macromolecular Modeling and Visualization, Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USADepartment of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Theoretical and Computational Biophysics Group, NIH Resource for Macromolecular Modeling and Visualization, Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USADepartment of Cell and Developmental Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USADepartment of Cell and Developmental Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USADepartment of Cell and Developmental Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USADepartment of Chemistry, Northwestern University, Evanston, IL, USADepartment of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Theoretical and Computational Biophysics Group, NIH Resource for Macromolecular Modeling and Visualization, Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Cancer Center at Illinois, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Department of Biomedical and Translational Sciences, Carle Illinois College of Medicine, University of Illinois at Urbana-Champaign, Urbana, IL, USADepartment of Cell and Developmental Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Cancer Center at Illinois, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Department of Biomedical and Translational Sciences, Carle Illinois College of Medicine, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Corresponding authorSummary: Although most members of the Rho guanine nucleotide exchange factor (RhoGEF) family are found to be phosphorylated, how phosphorylation regulates RhoGEF function is poorly understood. Here we report the discovery of a mechanism of RhoGEF regulation by phosphorylation. We find that ARHGEF3 is phosphorylated in the pleckstrin homology (PH) domain in a protein kinase C (PKC)-dependent manner. This phosphorylation inhibits ARHGEF3 activation of RhoA and actin stress fiber formation in the cells, and it also disrupts ARHGEF3 binding to PI(3,5)P2 but not to PI(4,5)P2. Guided by molecular dynamics simulation, a mutation in the PH domain is identified to uncouple ARHGEF3 binding to the two lipids, which is used to rule out a role of PI(3,5)P2 in regulating GEF activity. Results of in vitro GEF assays suggest that PH domain phosphorylation diminishes ARHGEF3 catalytic activity, likely through an allosteric mechanism. Our findings reveal a previously unknown type of regulatory mechanism for the family of RhoGEFs.http://www.sciencedirect.com/science/article/pii/S2589004225010144BiochemistryIn silico biologyMolecular biology |
| spellingShingle | Jesus F. Moreno Jae-Sung You Carlos C. Rodriguez Shashank Pant Adriana Reyes-Ordoñez Reean Abdullah Nilmani Singh Maxine J. van der Donk Emad Tajkhorshid Jie Chen Regulation of Rho guanine nucleotide exchange factor 3 by phosphorylation in the PH domain iScience Biochemistry In silico biology Molecular biology |
| title | Regulation of Rho guanine nucleotide exchange factor 3 by phosphorylation in the PH domain |
| title_full | Regulation of Rho guanine nucleotide exchange factor 3 by phosphorylation in the PH domain |
| title_fullStr | Regulation of Rho guanine nucleotide exchange factor 3 by phosphorylation in the PH domain |
| title_full_unstemmed | Regulation of Rho guanine nucleotide exchange factor 3 by phosphorylation in the PH domain |
| title_short | Regulation of Rho guanine nucleotide exchange factor 3 by phosphorylation in the PH domain |
| title_sort | regulation of rho guanine nucleotide exchange factor 3 by phosphorylation in the ph domain |
| topic | Biochemistry In silico biology Molecular biology |
| url | http://www.sciencedirect.com/science/article/pii/S2589004225010144 |
| work_keys_str_mv | AT jesusfmoreno regulationofrhoguaninenucleotideexchangefactor3byphosphorylationinthephdomain AT jaesungyou regulationofrhoguaninenucleotideexchangefactor3byphosphorylationinthephdomain AT carloscrodriguez regulationofrhoguaninenucleotideexchangefactor3byphosphorylationinthephdomain AT shashankpant regulationofrhoguaninenucleotideexchangefactor3byphosphorylationinthephdomain AT adrianareyesordonez regulationofrhoguaninenucleotideexchangefactor3byphosphorylationinthephdomain AT reeanabdullah regulationofrhoguaninenucleotideexchangefactor3byphosphorylationinthephdomain AT nilmanisingh regulationofrhoguaninenucleotideexchangefactor3byphosphorylationinthephdomain AT maxinejvanderdonk regulationofrhoguaninenucleotideexchangefactor3byphosphorylationinthephdomain AT emadtajkhorshid regulationofrhoguaninenucleotideexchangefactor3byphosphorylationinthephdomain AT jiechen regulationofrhoguaninenucleotideexchangefactor3byphosphorylationinthephdomain |