Bax- Bcl-xL interaction dynamics during the progression of cell cycle and cell death using FLIM-FRET
Genetically identical cells in a population show cell-to-cell variability because of fluctuation in transcription, epigenetics, post-translational modifications, and stochastic or extrinsically triggered non-genetic alterations. The change in the interaction state of proteins also emerges as an addi...
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| Format: | Article |
| Language: | English |
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Shared Science Publishers OG
2025-07-01
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| Series: | Cell Stress |
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| Online Access: | http://microbialcell.com/researcharticles/2025a-halikar-cell-stress |
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| author | Aman Munirpasha Halikar Aneesh Chandrasekharan Asha Lekshmi Aswathy Sivasailam Jain Tiffee P J Shivanshu Kumar Tiwari Aijaz Ahmad Rather TR Santhoshkumar |
| author_facet | Aman Munirpasha Halikar Aneesh Chandrasekharan Asha Lekshmi Aswathy Sivasailam Jain Tiffee P J Shivanshu Kumar Tiwari Aijaz Ahmad Rather TR Santhoshkumar |
| author_sort | Aman Munirpasha Halikar |
| collection | DOAJ |
| description | Genetically identical cells in a population show cell-to-cell variability because of fluctuation in transcription, epigenetics, post-translational modifications, and stochastic or extrinsically triggered non-genetic alterations. The change in the interaction state of proteins also emerges as an additional layer of cell signaling that influences cell cycle and cell death. However, the interrelation between cell cycle progression and cell death under the influence of spatio-temporal changes in protein-protein interaction is difficult to demonstrate in growing cells. This requires tools for cell cycle phase-resolved visualization of macromolecular interactions in live cells. We describe an approach to visualize the interaction of pro- and anti-death signaling partners, Bax and Bcl-xL, during cell cycle progression and cell death in live cells. Cells were stably expressed with Bax and Bcl-xL with FRET pairs and real-time cell cycle indicator probes. Acceptor photobleaching and Fluorescence lifetime imaging revealed interaction dynamics between Bax and Bcl-xL in isogenic stable cells. While Bcl-xL inhibited cell cycle progression, Bax promoted the cell cycle. The study highlighted an increased Bax-Bcl-xL interaction in the G1 phase compared to the non-G1 phase. Increased interaction is seen under stressed conditions and Bax-activated cells with FLIM-FRET, highlighting the nature of Bax-Bcl-xL interaction during cellular stress. In conclusion, our study explains Bax-Bcl-xL interaction dynamics in real-time and the potential utility of the approach to study macromolecular interactions along with cell cycle and cell death. |
| format | Article |
| id | doaj-art-40bbb8f90fa94fc6b77bef5a43cfdae5 |
| institution | Matheson Library |
| issn | 2523-0204 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Shared Science Publishers OG |
| record_format | Article |
| series | Cell Stress |
| spelling | doaj-art-40bbb8f90fa94fc6b77bef5a43cfdae52025-07-09T13:46:05ZengShared Science Publishers OGCell Stress2523-02042025-07-01914315710.15698/cst2025.07.307Bax- Bcl-xL interaction dynamics during the progression of cell cycle and cell death using FLIM-FRETAman Munirpasha Halikar0Aneesh Chandrasekharan1Asha Lekshmi2Aswathy Sivasailam3Jain Tiffee P J4Shivanshu Kumar Tiwari5Aijaz Ahmad Rather6TR Santhoshkumar7Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Poojappura, Thycaud P.O., Thiruvananthapuram, Kerala 695014, India.Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Poojappura, Thycaud P.O., Thiruvananthapuram, Kerala 695014, India.Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Poojappura, Thycaud P.O., Thiruvananthapuram, Kerala 695014, India.Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Poojappura, Thycaud P.O., Thiruvananthapuram, Kerala 695014, India.Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Poojappura, Thycaud P.O., Thiruvananthapuram, Kerala 695014, India.Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Poojappura, Thycaud P.O., Thiruvananthapuram, Kerala 695014, India.Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Poojappura, Thycaud P.O., Thiruvananthapuram, Kerala 695014, India.Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Poojappura, Thycaud P.O., Thiruvananthapuram, Kerala 695014, India.Genetically identical cells in a population show cell-to-cell variability because of fluctuation in transcription, epigenetics, post-translational modifications, and stochastic or extrinsically triggered non-genetic alterations. The change in the interaction state of proteins also emerges as an additional layer of cell signaling that influences cell cycle and cell death. However, the interrelation between cell cycle progression and cell death under the influence of spatio-temporal changes in protein-protein interaction is difficult to demonstrate in growing cells. This requires tools for cell cycle phase-resolved visualization of macromolecular interactions in live cells. We describe an approach to visualize the interaction of pro- and anti-death signaling partners, Bax and Bcl-xL, during cell cycle progression and cell death in live cells. Cells were stably expressed with Bax and Bcl-xL with FRET pairs and real-time cell cycle indicator probes. Acceptor photobleaching and Fluorescence lifetime imaging revealed interaction dynamics between Bax and Bcl-xL in isogenic stable cells. While Bcl-xL inhibited cell cycle progression, Bax promoted the cell cycle. The study highlighted an increased Bax-Bcl-xL interaction in the G1 phase compared to the non-G1 phase. Increased interaction is seen under stressed conditions and Bax-activated cells with FLIM-FRET, highlighting the nature of Bax-Bcl-xL interaction during cellular stress. In conclusion, our study explains Bax-Bcl-xL interaction dynamics in real-time and the potential utility of the approach to study macromolecular interactions along with cell cycle and cell death.http://microbialcell.com/researcharticles/2025a-halikar-cell-stressflim-fretprotein-protein interactionsapoptosisbaxbcl-xl |
| spellingShingle | Aman Munirpasha Halikar Aneesh Chandrasekharan Asha Lekshmi Aswathy Sivasailam Jain Tiffee P J Shivanshu Kumar Tiwari Aijaz Ahmad Rather TR Santhoshkumar Bax- Bcl-xL interaction dynamics during the progression of cell cycle and cell death using FLIM-FRET Cell Stress flim-fret protein-protein interactions apoptosis bax bcl-xl |
| title | Bax- Bcl-xL interaction dynamics during the progression of cell cycle and cell death using FLIM-FRET |
| title_full | Bax- Bcl-xL interaction dynamics during the progression of cell cycle and cell death using FLIM-FRET |
| title_fullStr | Bax- Bcl-xL interaction dynamics during the progression of cell cycle and cell death using FLIM-FRET |
| title_full_unstemmed | Bax- Bcl-xL interaction dynamics during the progression of cell cycle and cell death using FLIM-FRET |
| title_short | Bax- Bcl-xL interaction dynamics during the progression of cell cycle and cell death using FLIM-FRET |
| title_sort | bax bcl xl interaction dynamics during the progression of cell cycle and cell death using flim fret |
| topic | flim-fret protein-protein interactions apoptosis bax bcl-xl |
| url | http://microbialcell.com/researcharticles/2025a-halikar-cell-stress |
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