| Autor: |
Kaiser CE; College of Pharmacy, University of Arizona , Tucson, Arizona 85721, United States., Van Ert NA; College of Pharmacy, University of Arizona , Tucson, Arizona 85721, United States., Agrawal P; College of Pharmacy, University of Arizona , Tucson, Arizona 85721, United States., Chawla R; BIO5 Institute, University of Arizona , Tucson, Arizona 85721, United States., Yang D; College of Pharmacy, University of Arizona , Tucson, Arizona 85721, United States.; University of Arizona Cancer Center, University of Arizona , Tucson, Arizona 85724, United States.; BIO5 Institute, University of Arizona , Tucson, Arizona 85721, United States., Hurley LH; College of Pharmacy, University of Arizona , Tucson, Arizona 85721, United States.; University of Arizona Cancer Center, University of Arizona , Tucson, Arizona 85724, United States.; BIO5 Institute, University of Arizona , Tucson, Arizona 85721, United States. |
| Abstrakt: |
Activating KRAS mutations frequently occur in pancreatic, colorectal, and lung adenocarcinomas. While many attempts have been made to target oncogenic KRAS, no clinically useful therapies currently exist. Most efforts to target KRAS have focused on inhibiting the mutant protein; a less explored approach involves targeting KRAS at the transcriptional level. The promoter element of the KRAS gene contains a GC-rich nuclease hypersensitive site with three potential DNA secondary structure-forming regions. These are referred to as the Near-, Mid-, and Far-regions, on the basis of their proximity to the transcription start site. As a result of transcription-induced negative superhelicity, these regions can open up to form unique DNA secondary structures: G-quadruplexes on the G-rich strand and i-motifs on the C-rich strand. While the G-quadruplexes have been well characterized, the i-motifs have not been investigated as thoroughly. Here we show that the i-motif that forms in the C-rich Mid-region is the most stable and exists in a dynamic equilibrium with a hybrid i-motif/hairpin species and an unfolded hairpin species. The transcription factor heterogeneous nuclear ribonucleoprotein K (hnRNP K) was found to bind selectively to the i-motif species and to positively modulate KRAS transcription. Additionally, we identified a benzophenanthridine alkaloid that dissipates the hairpin species and destabilizes the interaction of hnRNP K with the Mid-region i-motif. This same compound stabilizes the three existing KRAS G-quadruplexes. The combined effect of the compound on the Mid-region i-motif and the G-quadruplexes leads to downregulation of KRAS gene expression. This dual i-motif/G-quadruplex-interactive compound presents a new mechanism to modulate gene expression. |
