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Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling
10:00

Assessment of Selective mRNA Translation in Mammalian Cells by Polysome Profiling

Published on: October 28, 2014

Translational repression of thymidylate synthase by targeting its mRNA

Divita Garg1, Alexander V Beribisky, Glauco Ponterini

  • 1Molecular and Cellular Modeling Group, Heidelberg Institute for Theoretical Studies (HITS), Schloss-Wolfsbrunnenweg 35, 69118 Heidelberg, Germany. Divita.Garg@tum.de

Nucleic Acids Research
|February 21, 2013
PubMed

Insights

Hoechst 33258 (HT) reduces cancer drug resistance by lowering thymidylate synthase (TS) protein levels. This compound targets TS mRNA translation, offering a new strategy against anti-cancer drug resistance.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Pharmacology

Background:

  • Drug resistance targeting thymidylate synthase (TS) is a significant obstacle in cancer therapy.
  • Overexpression of TS protein is a key factor contributing to this resistance.
  • Targeting TS mRNA translation offers a potential strategy to overcome drug resistance.

Purpose of the Study:

  • To investigate the effect of Hoechst 33258 (HT) on cellular TS protein levels.
  • To explore the mechanism by which HT modulates TS expression.
  • To elucidate the interaction between HT and TS mRNA.

Main Methods:

  • Nuclear magnetic resonance (NMR) spectroscopy
  • UV-visible and fluorescence spectroscopy
  • Molecular docking and dynamics simulations

Main Results:

  • HT significantly reduced cellular TS protein levels without affecting TS mRNA levels.
  • HT was found to interact with a specific region of TS mRNA near the translational initiation site.
  • The primary mode of interaction involved intercalation of HT at a CC mismatch within the TS mRNA.

Conclusions:

  • HT modulates TS expression at the translational level.
  • HT-like compounds show promise for developing novel therapeutic agents targeting TS mRNA.
  • This study provides a foundation for designing new anti-cancer drugs to combat TS-mediated resistance.

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