Single cell imaging reveals cisplatin regulating interactions between transcription (co)factors and DNA

期刊

Chemical Science

标题

Single cell imaging reveals cisplatin regulating interactions between transcription (co)factors and DNA

作者

Yu Lin, Kui Wu, Feifei Jia, Ling Chen, Zhaoying Wang, Yanyan Zhang, Qun Luo, Suyan Liu, Luyu Qi, Nan Li, Pu Dong, Fei Gao, Wei Zheng, Xiaohong Fang, Yao Zhao, Fuyi Wang摘要

Cisplatin is an extremely successful anticancer drug, and is commonly thought to target DNA. However, the way in which cisplatin-induced DNA lesions regulate interactions between transcription factors/cofactors and genomic DNA remains unclear. Herein, we developed a dual-modal microscopy imaging strategy to investigate, in situ, the formation of ternary binding complexes of the transcription cofactor HMGB1 and transcription factor Smad3 with cisplatin crosslinked DNA in single cells. We utilized confocal microscopy imaging to map EYFP-fused HMGB1 and fluorescent dye-stained DNA in single cells, followed by the visualization of cisplatin using high spatial resolution (200–350 nm) time of flight secondary ion mass spectrometry (ToF-SIMS) imaging of the same cells. The superposition of the fluorescence and the mass spectrometry (MS) signals indicate the formation of HMGB1-Pt-DNA ternary complexes in the cells. More significantly, for the first time, similar integrated imaging revealed that the cisplatin lesions at Smad-binding elements, for example GGC(GC)/(CG) and AGAC, disrupted the interactions of Smad3 with DNA, which was evidenced by the remarkable reduction in the expression of Smad-specific luciferase reporters subjected to cisplatin treatment. This finding suggests that Smad3 and its related signalling pathway are most likely involved in the intracellular response to cisplatin induced DNA damage.

原文链接

https://pubs.rsc.org/en/content/articlelanding/2021/sc/d0sc06760a

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