PUBLICATION
Subcellular drug targeting illuminates local kinase action
- Authors
- Bucko, P.J., Lombard, C.K., Rathbun, L., Garcia, I., Bhat, A., Wordeman, L., Smith, F.D., Maly, D.J., Hehnly, H., Scott, J.D.
- ID
- ZDB-PUB-191226-28
- Date
- 2019
- Source
- eLIFE 8: (Journal)
- Registered Authors
- Hehnly, Heidi
- Keywords
- biochemistry, cell biology, chemical biology, kinases, local signaling, mitosis, zebrafish
- MeSH Terms
-
- A Kinase Anchor Proteins/genetics*
- Animals
- Aurora Kinase A/chemistry
- Aurora Kinase A/genetics*
- Cell Cycle Proteins/chemistry
- Cell Cycle Proteins/genetics*
- Centrosome/chemistry
- Centrosome/ultrastructure
- Embryonic Development/genetics
- Gene Expression Regulation, Developmental
- Humans
- Kinetochores/chemistry
- Microtubules/genetics
- Mitosis/genetics*
- Phosphorylation/genetics
- Protein Kinase Inhibitors/chemistry
- Protein Serine-Threonine Kinases/chemistry
- Protein Serine-Threonine Kinases/genetics*
- Proto-Oncogene Proteins/chemistry
- Proto-Oncogene Proteins/genetics*
- Zebrafish/genetics
- Zebrafish/growth & development
- PubMed
- 31872801 Full text @ Elife
Citation
Bucko, P.J., Lombard, C.K., Rathbun, L., Garcia, I., Bhat, A., Wordeman, L., Smith, F.D., Maly, D.J., Hehnly, H., Scott, J.D. (2019) Subcellular drug targeting illuminates local kinase action. eLIFE. 8:.
Abstract
Deciphering how signaling enzymes operate within discrete microenvironments is fundamental to understanding biological processes. A-kinase anchoring proteins (AKAPs) restrict the range of action of protein kinases within intracellular compartments. We exploited the AKAP targeting concept to create genetically encoded platforms that restrain kinase inhibitor drugs at distinct subcellular locations. Local Kinase Inhibition (LoKI) allows us to ascribe organelle-specific functions to broad specificity kinases. Using chemical genetics, super resolution microscopy, and live-cell imaging we discover that centrosomal delivery of Polo-like kinase 1 (Plk1) and Aurora A (AurA) inhibitors attenuates kinase activity, produces spindle defects, and prolongs mitosis. Targeted inhibition of Plk1 in zebrafish embryos illustrates how centrosomal Plk1 underlies mitotic spindle assembly. Inhibition of kinetochore-associated pools of AurA blocks phosphorylation of microtubule-kinetochore components. This versatile precision pharmacology tool enhances investigation of local kinase biology.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping