<i>O</i>-GlcNAcylation of Focal Adhesion Kinase Regulates Cell Adhesion, Migration, and Proliferation via the FAK/AKT Pathway
Focal Adhesion Kinase (FAK) is a non-receptor tyrosine kinase pivotal in cellular signal transduction, regulating cell adhesion, migration, growth, and survival. However, the regulatory mechanisms of FAK during tumorigenesis and progression still need to be fully understood. Our previous study demon...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-12-01
|
| Series: | Biomolecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2218-273X/14/12/1577 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850086407448559616 |
|---|---|
| author | Zhiwei Zhang Tomoya Isaji Yoshiyuki Oyama Jianwei Liu Zhiwei Xu Yuhan Sun Tomohiko Fukuda Haojie Lu Jianguo Gu |
| author_facet | Zhiwei Zhang Tomoya Isaji Yoshiyuki Oyama Jianwei Liu Zhiwei Xu Yuhan Sun Tomohiko Fukuda Haojie Lu Jianguo Gu |
| author_sort | Zhiwei Zhang |
| collection | DOAJ |
| description | Focal Adhesion Kinase (FAK) is a non-receptor tyrosine kinase pivotal in cellular signal transduction, regulating cell adhesion, migration, growth, and survival. However, the regulatory mechanisms of FAK during tumorigenesis and progression still need to be fully understood. Our previous study demonstrated that <i>O</i>-GlcNAcylation regulates integrin-mediated cell adhesion. To further elucidate the underlying molecular mechanism, we focused on FAK in this study and purified it from 293T cells. Using liquid chromatography–mass spectrometry (LC-MS/MS), we identified the <i>O</i>-GlcNAcylation of FAK at Ser708, Thr739, and Ser886. Compared with wild-type FAK expressed in <i>FAK</i>-knockout 293T cells, the FAK mutant, in which Ser708, Thr739, and Ser886 were replaced with Ala, exhibited lower phosphorylation levels of Tyr397 and AKT. Cell proliferation and migration, assessed through MTT and wound healing assays, were significantly suppressed in the FAK mutant cells compared to the wild-type FAK cells. Additionally, the interaction among FAK, paxillin, and talin was enhanced, and cell adhesion was increased in the mutant cells. These data indicate that specific <i>O</i>-GlcNAcylation of FAK plays a critical regulatory role in integrin-mediated cell adhesion and migration. This further supports the idea that <i>O</i>-GlcNAcylation is essential for tumorigenesis and progression and that targeting the <i>O</i>-GlcNAcylation of FAK could offer a promising therapeutic strategy for cancer treatment. |
| format | Article |
| id | doaj-art-48bd61905fc34d5c864e9686e994ca4b |
| institution | DOAJ |
| issn | 2218-273X |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj-art-48bd61905fc34d5c864e9686e994ca4b2025-08-20T02:43:29ZengMDPI AGBiomolecules2218-273X2024-12-011412157710.3390/biom14121577<i>O</i>-GlcNAcylation of Focal Adhesion Kinase Regulates Cell Adhesion, Migration, and Proliferation via the FAK/AKT PathwayZhiwei Zhang0Tomoya Isaji1Yoshiyuki Oyama2Jianwei Liu3Zhiwei Xu4Yuhan Sun5Tomohiko Fukuda6Haojie Lu7Jianguo Gu8Division of Regulatory Glycobiology, Graduate School of Pharmaceutical Sciences, Tohoku Medical and Pharmaceutical University, Sendai 980-0845, Miyagi, JapanDivision of Regulatory Glycobiology, Graduate School of Pharmaceutical Sciences, Tohoku Medical and Pharmaceutical University, Sendai 980-0845, Miyagi, JapanDivision of Regulatory Glycobiology, Graduate School of Pharmaceutical Sciences, Tohoku Medical and Pharmaceutical University, Sendai 980-0845, Miyagi, JapanDivision of Regulatory Glycobiology, Graduate School of Pharmaceutical Sciences, Tohoku Medical and Pharmaceutical University, Sendai 980-0845, Miyagi, JapanDivision of Regulatory Glycobiology, Graduate School of Pharmaceutical Sciences, Tohoku Medical and Pharmaceutical University, Sendai 980-0845, Miyagi, JapanDivision of Regulatory Glycobiology, Graduate School of Pharmaceutical Sciences, Tohoku Medical and Pharmaceutical University, Sendai 980-0845, Miyagi, JapanDivision of Regulatory Glycobiology, Graduate School of Pharmaceutical Sciences, Tohoku Medical and Pharmaceutical University, Sendai 980-0845, Miyagi, JapanShanghai Cancer Center and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, ChinaDivision of Regulatory Glycobiology, Graduate School of Pharmaceutical Sciences, Tohoku Medical and Pharmaceutical University, Sendai 980-0845, Miyagi, JapanFocal Adhesion Kinase (FAK) is a non-receptor tyrosine kinase pivotal in cellular signal transduction, regulating cell adhesion, migration, growth, and survival. However, the regulatory mechanisms of FAK during tumorigenesis and progression still need to be fully understood. Our previous study demonstrated that <i>O</i>-GlcNAcylation regulates integrin-mediated cell adhesion. To further elucidate the underlying molecular mechanism, we focused on FAK in this study and purified it from 293T cells. Using liquid chromatography–mass spectrometry (LC-MS/MS), we identified the <i>O</i>-GlcNAcylation of FAK at Ser708, Thr739, and Ser886. Compared with wild-type FAK expressed in <i>FAK</i>-knockout 293T cells, the FAK mutant, in which Ser708, Thr739, and Ser886 were replaced with Ala, exhibited lower phosphorylation levels of Tyr397 and AKT. Cell proliferation and migration, assessed through MTT and wound healing assays, were significantly suppressed in the FAK mutant cells compared to the wild-type FAK cells. Additionally, the interaction among FAK, paxillin, and talin was enhanced, and cell adhesion was increased in the mutant cells. These data indicate that specific <i>O</i>-GlcNAcylation of FAK plays a critical regulatory role in integrin-mediated cell adhesion and migration. This further supports the idea that <i>O</i>-GlcNAcylation is essential for tumorigenesis and progression and that targeting the <i>O</i>-GlcNAcylation of FAK could offer a promising therapeutic strategy for cancer treatment.https://www.mdpi.com/2218-273X/14/12/1577cell adhesioncellular signalingfocal adhesion kinase<i>O</i>-GlcNAcylationOGT |
| spellingShingle | Zhiwei Zhang Tomoya Isaji Yoshiyuki Oyama Jianwei Liu Zhiwei Xu Yuhan Sun Tomohiko Fukuda Haojie Lu Jianguo Gu <i>O</i>-GlcNAcylation of Focal Adhesion Kinase Regulates Cell Adhesion, Migration, and Proliferation via the FAK/AKT Pathway Biomolecules cell adhesion cellular signaling focal adhesion kinase <i>O</i>-GlcNAcylation OGT |
| title | <i>O</i>-GlcNAcylation of Focal Adhesion Kinase Regulates Cell Adhesion, Migration, and Proliferation via the FAK/AKT Pathway |
| title_full | <i>O</i>-GlcNAcylation of Focal Adhesion Kinase Regulates Cell Adhesion, Migration, and Proliferation via the FAK/AKT Pathway |
| title_fullStr | <i>O</i>-GlcNAcylation of Focal Adhesion Kinase Regulates Cell Adhesion, Migration, and Proliferation via the FAK/AKT Pathway |
| title_full_unstemmed | <i>O</i>-GlcNAcylation of Focal Adhesion Kinase Regulates Cell Adhesion, Migration, and Proliferation via the FAK/AKT Pathway |
| title_short | <i>O</i>-GlcNAcylation of Focal Adhesion Kinase Regulates Cell Adhesion, Migration, and Proliferation via the FAK/AKT Pathway |
| title_sort | i o i glcnacylation of focal adhesion kinase regulates cell adhesion migration and proliferation via the fak akt pathway |
| topic | cell adhesion cellular signaling focal adhesion kinase <i>O</i>-GlcNAcylation OGT |
| url | https://www.mdpi.com/2218-273X/14/12/1577 |
| work_keys_str_mv | AT zhiweizhang ioiglcnacylationoffocaladhesionkinaseregulatescelladhesionmigrationandproliferationviathefakaktpathway AT tomoyaisaji ioiglcnacylationoffocaladhesionkinaseregulatescelladhesionmigrationandproliferationviathefakaktpathway AT yoshiyukioyama ioiglcnacylationoffocaladhesionkinaseregulatescelladhesionmigrationandproliferationviathefakaktpathway AT jianweiliu ioiglcnacylationoffocaladhesionkinaseregulatescelladhesionmigrationandproliferationviathefakaktpathway AT zhiweixu ioiglcnacylationoffocaladhesionkinaseregulatescelladhesionmigrationandproliferationviathefakaktpathway AT yuhansun ioiglcnacylationoffocaladhesionkinaseregulatescelladhesionmigrationandproliferationviathefakaktpathway AT tomohikofukuda ioiglcnacylationoffocaladhesionkinaseregulatescelladhesionmigrationandproliferationviathefakaktpathway AT haojielu ioiglcnacylationoffocaladhesionkinaseregulatescelladhesionmigrationandproliferationviathefakaktpathway AT jianguogu ioiglcnacylationoffocaladhesionkinaseregulatescelladhesionmigrationandproliferationviathefakaktpathway |