c-Abl Tyrosine Kinase Is Regulated Downstream of the Cytoskeletal Protein Synemin in Head and Neck Squamous Cell Carcinoma Radioresistance and DNA Repair
| Autor: | Anne Vehlow, Nils Cordes, Sara Sofia Deville, Luis F. Delgadillo Silva |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Embryo
Nonmammalian DNA Repair Cell Culture Techniques Ataxia Telangiectasia Mutated Proteins SH2 domain HNSCC Radiation Tolerance lcsh:Chemistry 0302 clinical medicine Intermediate Filament Proteins hemic and lymphatic diseases DNA Breaks Double-Stranded synemin Phosphorylation RNA Small Interfering Tyrosine Proto-Oncogene Proteins c-abl lcsh:QH301-705.5 Zebrafish Spectroscopy 0303 health sciences Synemin Chemistry DNA Neoplasm General Medicine Computer Science Applications Cell biology Gene Expression Regulation Neoplastic ionizing radiation c-Abl DNA repair zebrafish Head and Neck Neoplasms 030220 oncology & carcinogenesis Tyrosine kinase Protein Binding Signal Transduction Ionizing radiation Article Catalysis Inorganic Chemistry 03 medical and health sciences Cell Line Tumor Radioresistance Animals Humans Protein Interaction Domains and Motifs Physical and Theoretical Chemistry Kinase activity Molecular Biology Cell Proliferation 030304 developmental biology Squamous Cell Carcinoma of Head and Neck X-Rays Organic Chemistry lcsh:Biology (General) lcsh:QD1-999 Cancer cell |
| Zdroj: | International Journal of Molecular Sciences International Journal of Molecular Sciences; Volume 21; Issue 19; Pages: 7277 International Journal of Molecular Sciences, Vol 21, Iss 7277, p 7277 (2020) International Journal of Molecular Sciences 21(2020)19, 7277 |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms21197277 |
| Popis: | The intermediate filament synemin has been previously identified as novel regulator of cancer cell therapy resistance and DNA double strand break (DSB) repair. c-Abl tyrosine kinase is involved in both of these processes. Using PamGene technology, we performed a broad-spectrum kinase activity profiling in three-dimensionally, extracellular matrix grown head and neck cancer cell cultures. Upon synemin silencing, we identified 86 deactivated tyrosine kinases, including c-Abl, in irradiated HNSCC cells. Upon irradiation and synemin inhibition, c-Abl hyperphosphorylation on tyrosine (Y) 412 and threonine (T) 735 was significantly reduced, prompting us to hypothesize that c-Abl tyrosine kinase is an important signaling component of the synemin-mediated radioresistance pathway. Simultaneous targeting of synemin and c-Abl resulted in similar radiosensitization and DSB repair compared with single synemin depletion, suggesting synemin as an upstream regulator of c-Abl. Immunoprecipitation assays revealed a protein complex formation between synemin and c-Abl pre- and post-irradiation. Upon pharmacological inhibition of ATM, synemin/c-Abl protein-protein interactions were disrupted implying synemin function to depend on ATM kinase activity. Moreover, deletion of the SH2 domain of c-Abl demonstrated a decrease in interaction, indicating the dependency of the protein-protein interaction on this domain. Mechanistically, radiosensitization upon synemin knockdown seems to be associated with an impairment of DNA repair via regulation of non-homologous end joining independent of c-Abl function. Our data generated in more physiological 3D cancer cell culture models suggest c-Abl as further key determinant of radioresistance downstream of synemin. |
| Databáze: | OpenAIRE |
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