Morphological effects of g-quadruplex stabilization using a small molecule in zebrafish
- Authors
- Agarwal, T., Lalwani, M.K., Kumar, S., Roy, S., Chakraborty, T.K., Sivasubbu, S., and Maiti, S.
- ID
- ZDB-PUB-140402-21
- Date
- 2014
- Source
- Biochemistry 53(7): 1117-1124 (Journal)
- Registered Authors
- Sivasubbu, Sridhar
- Keywords
- none
- MeSH Terms
-
- Animals
- Cadherins/genetics*
- Calorimetry
- Circular Dichroism
- G-Quadruplexes/drug effects*
- Ligands
- Molecular Structure
- Nucleotide Motifs
- Peptides, Cyclic/chemistry*
- Peptides, Cyclic/pharmacology*
- Promoter Regions, Genetic/genetics
- Structure-Activity Relationship
- Zebrafish*/embryology
- PubMed
- 24476096 Full text @ Biochemistry
Zebrafish (Danio rerio) embryos are transparent and advantageous for studying early developmental changes due to ex utero development, making them an appropriate model for studying gene expression changes as a result of molecular targeting. Zebrafish embryos were injected with a previously reported G-quadruplex selective ligand, and the phenotypic changes were recorded. We report marked discrepancies in the development of intersegmental vessels. In silico analysis determined that the putative G-quadruplex motif occur in the upstream promoter region of the Cdh5 (N-cadherin) gene. A real-time polymerase chain reaction-based investigation indicated that in zebrafish, CDH-2 (ZN-cad) was significantly downregulated in the ligand-treated embryos. Biophysical characterization of the interaction of the ligand with the G-quadruplex motif found in this promoter yielded strong binding and stabilization of the G-quadruplex with this ligand. Hence, we report for the first time the phenotypic impact of G-quadruplex targeting with a ligand in a vertebrate organism. This study has unveiled not only G-quadruplex targeting in non-human animal species but also the potential that G-quadruplexes can provide a ready tool for understanding the phenotypic effects of targeting certain important genes involved in differentiation and developmental processes in a living eukaryotic organism.