Orginal Article |
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Identification and Functional Study of G-quadruplexes in SENP1 Promoter |
ZHOU Yan-xing,HAN Meng,LIU Na-nv,HUANG Wei-wei() |
College of Life Science, Northwest A & F University, Yangling 712100, China |
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Abstract Objective: The regulation effect of G-quadruplexes (G4) on the SUMO-specific proteases 1 (SENP1) transcriptional expression was studied. Method: Reporting plasmids of SENP1 promoter were constructed by cloning different SENP1 promoter fragments. The core transcriptional regulatory region of SENP1 promoter was identified by using reporter assay. The sequence of core transcriptional regulatory region of SENP1 promoter was analyzed and G-quadruplex (G4) formation sequence was predicted. Oligonucleotides of G4 formation sequence from SENP1 promoter were synthesized, and their topological structure was detected by circular dichroism analysis. G4 ligand TMPyP4 and G4 helicase G4R1 were used to detect the regulatory effect of promoter G4 on SENP1 transcriptional expression by using reporter assay and Western blot. Result: The -910~+226 region was found to be the core transcriptional regulatory region of SENP1 promoter. Sequence analysis showed the core region of SENP1 promoter is rich in G/C and contains G4 formation sequences. Circular dichroism analysis confirmed that the oligonucleotides of G4 formation sequence from SENP1 promoter form G4 structure. Reporter assay and Western blot showed that promoter G4 inhibited the transcriptional expression of SENP1. Conclusion: G4 exist in the core transcriptional regulatory region of SENP1 promoter and show negative regulation on SENP1 transcriptional expression, which provides new research ideas and experimental clues for revealing the mechanism of SENP1 in physiological and pathological processes.
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Received: 05 May 2019
Published: 27 March 2020
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Corresponding Authors:
Wei-wei HUANG
E-mail: whuang0210@163.com
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