Citation: CHANG Tian-Jun, WU Li, BIAN Wen-Xiu, BING Tao, ZHAO Tong-Qian, SHANGGUAN Di-Hua. In-Gel Probing Polymorphic Structures of G-Quadruplexes Derived From c-Myc Promoter[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(3): 323-331. doi: 10.19756/j.issn.0253-3820.191603 shu

In-Gel Probing Polymorphic Structures of G-Quadruplexes Derived From c-Myc Promoter

1.
Department of Biology, Institute of Resources and Environment, Henan Polytechnic University, Jiaozuo 454000, China;
2.
Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China

Received Date: 13 October 2019
Revised Date: 20 December 2019

Fund Project: This work was supported by the National Natural Science Foundation of China (No. U1704241), the Plan for Scientific Innovation Talent of Henan Province, China (No. 194200510010), and the Natural Science Foundation of Henan Province, China (No. 162300410111).

Some G-quadruplex (G4) forming sequences are able to form multimers under physiological conditions, which limits the structure-activity study of G4s. Here we reported an in-gel visual method to probe the conformational polymorphism of the derivatives of a parallel G4-forming sequence, c-Myc by flanking with different A/T bases and typical intermolecular G4-forming sequences. Fluorescence and circular dichroism (CD) spectra showed that the interaction of c-Myc derivatives with Thioflavin T (ThT) caused different fluorescence enhancements of ThT, and intermolecular parallel G4s caused higher fluorescence enhancement of ThT than that of intramolecular parallel and intermolecular antiparallel G4s. These DNA sequences were separated on native gels and stained by ThT for G4 and then by Stains-all (dye) for all DNA structures. Most of the sequences adopted parallel G4 multimers and intramolecular G4s in the presence of K⁺, meanwhile only a part of them formed G4 multimers and intramolecular structures in the presence of Na⁺. The optical density (OD) ratio of DNA bands stained by ThT and Stains-all were derived by ImageJ, and the ratios of G4 multimers were higher than 1.0, intramolecular parallel G4s were about 0.5, and non-G4s were near 0. Furthermore, the flanking nucleotides were favorable to the formation of multimers of c-Myc in K⁺ solutions, especially those flanking with T; while in Na⁺ solutions, only those sequences flanking with two or more A preferred to form G4 multimers. The results suggested the current strategy had the potential for the polymorphic study of G4s.
- G-quadruplex,
- In situ detection,
- Structural polymorphism,
- Molecular recognition,
- Fluorescence probe
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沈阳化工大学材料科学与工程学院沈阳 110142