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

  • Tuom T.T Truong
  • Thu M.T. Dao
  • Trang P.T Phan
  • Hoang D Nguyen
  • Dung H Nguyen
  • Dung Thanh Dang
Từ khóa: G4-RHAU interaction, Inhibition, Protein expression

Tóm tắt

Introduction: 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.

Methods: A sequence 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.

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