Identification and characterization of G-quadruplex formation within the EP0 promoter of pseudorabies virus

被引:21
作者
Kong, Jiang-Nan [1 ]
Zhang, Chao [1 ]
Zhu, Yan-Ce [1 ]
Zhong, Kai [1 ]
Wang, Jiang [1 ]
Chu, Bei-Bei [1 ]
Yang, Guo-Yu [1 ]
机构
[1] Henan Agr Univ, Coll Anim Sci & Vet Med, Zhengzhou 450002, Henan, Peoples R China
来源
SCIENTIFIC REPORTS | 2018年 / 8卷
基金
中国国家自然科学基金;
关键词
TRANSCRIPTION FACTOR; CIRCULAR-DICHROISM; TELOMERIC DNA; SMALL-MOLECULE; PROTEIN; REGION; BINDING; GENE; RNA; INHIBITION;
D O I
10.1038/s41598-018-32222-7
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
EP0 is an important early gene that modulates the life cycle of pseudorabies virus (PRV). A guaninerich sequence overlapping with three Sp1 binding sites is located upstream of the transcription start site (TSS) in the EP0 promoter. Using native polyacrylamide gel electrophoresis (PAGE) and circular dichroism (CD), we verified that the G-rich region in the EP0 promoter forms an intramolecular parallel G-quadruplex (G4) in the presence of K+ ions. Further dimethyl sulphate (DMS) footprinting and Taq polymerase stop assays indicates the potential polymorphic folding of G4. In addition, a small chemical ligand, pyridostatin (PDS), promotes and stabilizes the formation of G4. Interestingly, based on the results of electrophoretic mobility shift assays (EMSA), the Sp1 protein bound to G4-bearing DNA with more affinity than DNA lacking the G4 structure. According to the luciferase reporter assay, G4 negatively regulates the EP0 promoter activity. These results demonstrate that Sp1 and G4 cooperate to regulate EP0 promoter activity.
引用
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页数:13
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