PUBLICATION
Evolutionarily conserved intercalated disc protein Tmem65 regulates cardiac conduction and connexin 43 function
- Authors
- Sharma, P., Abbasi, C., Lazic, S., Teng, A.C., Wang, D., Dubois, N., Ignatchenko, V., Wong, V., Liu, J., Araki, T., Tiburcy, M., Ackerley, C., Zimmermann, W.H., Hamilton, R., Sun, Y., Liu, P.P., Keller, G., Stagljar, I., Scott, I.C., Kislinger, T., Gramolini, A.O.
- ID
- ZDB-PUB-150926-8
- Date
- 2015
- Source
- Nature communications 6: 8391 (Journal)
- Registered Authors
- Lazic, Savo, Scott, Ian
- Keywords
- Arrhythmias, Cell signalling, Heart development, Membrane proteins
- MeSH Terms
-
- Animals
- Blotting, Western
- Chromatography, High Pressure Liquid
- Connexin 43/metabolism*
- Fluorescent Antibody Technique
- Gap Junctions/metabolism*
- Gap Junctions/ultrastructure
- Gene Knockdown Techniques
- Heart Conduction System/metabolism*
- Heart Conduction System/physiology
- Heart Ventricles/metabolism*
- In Vitro Techniques
- Membrane Proteins/genetics*
- Membrane Proteins/metabolism
- Mice
- Microscopy, Electron
- Myocytes, Cardiac/metabolism*
- Myocytes, Cardiac/physiology
- Myocytes, Cardiac/ultrastructure
- Proteomics
- Silicon Dioxide
- Zebrafish
- Zebrafish Proteins/genetics*
- Zebrafish Proteins/metabolism
- PubMed
- 26403541 Full text @ Nat. Commun.
Citation
Sharma, P., Abbasi, C., Lazic, S., Teng, A.C., Wang, D., Dubois, N., Ignatchenko, V., Wong, V., Liu, J., Araki, T., Tiburcy, M., Ackerley, C., Zimmermann, W.H., Hamilton, R., Sun, Y., Liu, P.P., Keller, G., Stagljar, I., Scott, I.C., Kislinger, T., Gramolini, A.O. (2015) Evolutionarily conserved intercalated disc protein Tmem65 regulates cardiac conduction and connexin 43 function. Nature communications. 6:8391.
Abstract
Membrane proteins are crucial to heart function and development. Here we combine cationic silica-bead coating with shotgun proteomics to enrich for and identify plasma membrane-associated proteins from primary mouse neonatal and human fetal ventricular cardiomyocytes. We identify Tmem65 as a cardiac-enriched, intercalated disc protein that increases during development in both mouse and human hearts. Functional analysis of Tmem65 both in vitro using lentiviral shRNA-mediated knockdown in mouse cardiomyocytes and in vivo using morpholino-based knockdown in zebrafish show marked alterations in gap junction function and cardiac morphology. Molecular analyses suggest that Tmem65 interaction with connexin 43 (Cx43) is required for correct localization of Cx43 to the intercalated disc, since Tmem65 deletion results in marked internalization of Cx43, a shorter half-life through increased degradation, and loss of Cx43 function. Our data demonstrate that the membrane protein Tmem65 is an intercalated disc protein that interacts with and functionally regulates ventricular Cx43.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping