Inhibition of protein expression by the interaction of G-Quadruplex and RHAU peptide in E. coli

G-quadurplex (G4) formation plays a role in many biological processes such as replication, transcription, translation, and telomeric maintenance. Stabilization of G4 structure by peptide has recently emerged as a potential approach in the regulation of protein expression. In this study, we reported on regulation of cyan fluorescent protein (CFP) expression by the interaction of G4 and RNA helicase associated with AU-rich elements (RHAU) peptide in E. coli.
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Inhibition of protein expression by the interaction of G-Quadruplex and RHAU peptide in E. coliScience & Technology Development Journal, 22(4):378-384 Open Access Full Text Article Original ResearchInhibition of protein expression by the interaction ofG-Quadruplex and RHAU peptide in E. coliTuom T.T Truong1 , Thu M.T. Dao1 , Trang P.T Phan1 , Hoang D Nguyen1 , Dung H Nguyen2 , Dung T Dang1,3,* ABSTRACT Introduction: G-quadurplex (G4) formation plays a role in many biological processes such as repli- cation, transcription, translation, and telomeric maintenance. Stabilization of G4 structure by pep-Use your smartphone to scan this tide has recently emerged as a potential approach in the regulation of protein expression. In thisQR code and download this article study, we reported on regulation of cyan fluorescent protein (CFP) expression by the interaction of G4 and RNA helicase associated with AU-rich elements (RHAU) peptide in E. coli. Methods: A se- quence of TTGGGTGGGTGGGTGGGT (formed into G4 structure) was genetically applied to cfp gene as a reporter gene (g4-cfp). Both g4-cfp and DHX36 (or ∆DHX36) genes were cloned to pET-Duet1 vector that allowed to simultaneously express both G4-CFP protein and RHAU (or ∆RHAU) protein under IPTG inducer. Results: The level of G4-CFP expression in the presence of RHAU (pD64) was around 2-fold and 3-fold lower than that of G4-CFP expression in the presence of ∆RHAU (pD65) and G4-CFP expression alone (pD21), respectively. Conclusion: RHAU might selectively bind G4 structure of mRNA of G4-CFP, resulting in inhibition of G4-CFP expression in E. coli. The G4 and RHAU peptide interaction would provide a promising approach for inhibition of gene expression in many biomedical applications. Key words: G4-RHAU interaction, Inhibition, Protein expression1Center for Bioscience andBiotechnology, University of Science,National University-HCMC, Vietnam INTRODUCTION G4 structure in the presence of ATP in the cells. How- ever, only N-region of RHAU peptide (without heli-2Institue of Tropical Biology, VAST G-quadruplexes (G4s) are G-rich sequences that can case domain) can specifically bind and stabilize par-3Faculty of Biotechnology, Ho Chi Minh fold into four single-stranded DNA or RNA structures allel G4 structure 13 . Insight into the structure of G4City Open University, Vietnam by hoogsteen hydrogen interaction 1 (Figure 1). G4 structures can be parallel or nonparallel topology 2 . and RHAU peptide showed the RHAU covers a termi-Correspondence nal guanine base and binds the parallel G4 using 3 an- In the cell, the formation of G4 structure plays a cru-Dung T Dang, Center for Bioscience and chor point electrostatic interactions between negativeBiotechnology, University of Science, cial role in many biological processes such as replica-National University-HCMC, Vietnam tion, transcription, translation and telomeric mainte- charge of phosphate groups and 3 positive charges ofFaculty of Biotechnology, Ho Chi Minh nance 3 . For instance, the formation of G4 in the un- amino acids 14 . The studies of the interaction of G4City Open University, Vietnam translated region (UTR) that significantly affected the and RHAU peptide for biological applications haveEmail: dung.dthanh@ou.edu.vn level expression of protein 4–6 . Formation of G4 struc- also been reported. Incorporating RHAU peptide toHistory ture at the telomeric region that can prevent binding CFP allowed to generate the fluorescent probe which• Received: 2019-10-01 of telomerase to its DNA target leads to inhibition of can visualize and distinguish G4 topologies 15 . De-• Accepted: 2019-12-24 velopment of the novel ribonuclease by fusing RHAU telomeric elongation 7–9 . Therefore, the formation of• Published: 2019-12-31 G4s in DNA or RNA is considered to be a new molec- with RNase H catalytic domain that can selectivelyDOI : 10.32508/stdj.v22i4.1712 bind G4 and cleave RNA in RNA:DNA hybrid at the ular target for molecules in cancer therapeutics 10–12 . Specific recognition and stabilization of G4 by pep- specific manner 16 . However, the application of stabi- ...