PUBLICATION
A Chemical Biology Approach to Model Pontocerebellar Hypoplasia Type 1B (PCH1B)
- Authors
- Francois-Moutal, L., Jahanbakhsh, S., Nelson, A., Ray, D., Scott, D.D., Hennefarth, M., Moutal, A., Perez-Miller, S., Ambrose, A.J., Al-Shamari, A., Coursodon, P., Meechoovet, B., Reiman, R., Lyons, E., Beilstein, M., Chapman, E., Morris, Q.D., Van Keuren-Jensen, K., Hughes, T.R., Khanna, R., Koehler, C., Jen, J., Gokhale, V., Khanna, M.
- ID
- ZDB-PUB-180825-7
- Date
- 2018
- Source
- ACS Chemical Biology 13(10): 3000-3010 (Journal)
- Registered Authors
- Koehler, Carla
- Keywords
- none
- MeSH Terms
-
- Animals
- Atrophy
- Cerebellum/pathology
- Disease Models, Animal*
- Down-Regulation
- Exosome Multienzyme Ribonuclease Complex/chemistry
- Exosome Multienzyme Ribonuclease Complex/genetics
- Exosome Multienzyme Ribonuclease Complex/metabolism*
- Gene Knockdown Techniques
- Humans
- Isoquinolines/metabolism
- Isoquinolines/pharmacology*
- Isoquinolines/toxicity*
- Molecular Docking Simulation
- Mutation
- Olivopontocerebellar Atrophies/chemically induced
- Olivopontocerebellar Atrophies/genetics*
- Olivopontocerebellar Atrophies/pathology
- Phenotype
- Protein Binding
- Protein Domains
- RNA/metabolism*
- RNA-Binding Proteins/chemistry
- RNA-Binding Proteins/genetics
- RNA-Binding Proteins/metabolism*
- Spinal Curvatures/chemically induced
- Transcriptome/drug effects
- Up-Regulation
- Zebrafish/abnormalities*
- PubMed
- 30141626 Full text @ ACS Chem. Biol.
Citation
Francois-Moutal, L., Jahanbakhsh, S., Nelson, A., Ray, D., Scott, D.D., Hennefarth, M., Moutal, A., Perez-Miller, S., Ambrose, A.J., Al-Shamari, A., Coursodon, P., Meechoovet, B., Reiman, R., Lyons, E., Beilstein, M., Chapman, E., Morris, Q.D., Van Keuren-Jensen, K., Hughes, T.R., Khanna, R., Koehler, C., Jen, J., Gokhale, V., Khanna, M. (2018) A Chemical Biology Approach to Model Pontocerebellar Hypoplasia Type 1B (PCH1B). ACS Chemical Biology. 13(10):3000-3010.
Abstract
Mutations of EXOSC3 have been linked to the rare neurological disorder known as Pontocerebellar Hypoplasia type 1B (PCH1B). EXOSC3 is one of three putative RNA-binding structural cap proteins that guide RNA into the RNA exosome, the cellular machinery that degrades RNA. Using RNAcompete, we identified a G-rich RNA motif binding to EXOSC3. Surface Plasmon Resonance (SPR) and Microscale Thermophoresis (MST) indicated an affinity in the low micromolar range of EXOSC3 for long and short G-rich RNA sequences. Although several PCH1B-causing mutations in EXOSC3 did not engage a specific RNA motif as shown by RNA compete they exhibited lower binding affinity to G-rich RNA as demonstrated by MST. To test the hypothesis that modification of the RNA-protein interface in EXOSC3 mutants may be phenocopied by small molecules, we performed an in-silico screen of 50,000 small molecules and used enzyme-linked immunosorbant assays (ELISAs) and MST to assess the ability of the molecules to inhibit RNA-binding by EXOSC3. We identified a small molecule, EXOSC3-RNA disrupting (ERD) compound 3 (ERD03), which: (i) bound specifically to EXOSC3 in saturation transfer difference nuclear magnetic resonance (STD NMR); (ii) disrupted the EXOSC3-RNA interaction in a concentration-dependent manner; (iii) produced a PCH1B-like phenotype with a 50% reduction in the cerebellum and an abnormally curved spine in zebrafish embryos. This compound also induced modification of zebrafish RNA expression levels similar to that observed with a morpholino against EXOSC3. To our knowledge, this is the first example of a small molecule obtained by rational design that models the abnormal developmental effects of a neurodegenerative disease in a whole organism.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping