Alternative splicing controls myotonic dystrophy protein kinase structure, enzymatic activity, and subcellular localization

Autor: M. Coerwinkel-Driessen, Derick G. Wansink, Patricia J. T. A. Groenen, Bé Wieringa, René E. M. A. van Herpen, Brian A. Hemmings
Rok vydání: 2003
Předmět:
Gene isoform
DNA
Complementary
Protein Conformation
Amino Acid Motifs
Blotting
Western
Molecular Sequence Data
Protein Serine-Threonine Kinases
Biology
Endoplasmic Reticulum
Models
Biological
Myotonic dystrophy
Myotonin-Protein Kinase
Mice
Myosin-Light-Chain Phosphatase
Cytosol
Cell Movement
Phosphoprotein Phosphatases
medicine
Animals
Protein Isoforms
Amino Acid Sequence
Phosphorylation
Protein kinase A
Cell Growth and Development
Molecular Biology
Rho-associated protein kinase
Models
Genetic
Sequence Homology
Amino Acid
Kinase
Autophosphorylation
Alternative splicing
Cell Biology
Subcellular localization
medicine.disease
Precipitin Tests
Neuromuscular development and genetic disorders [UMCN 3.1]
Mitochondria
Protein Structure
Tertiary
Tumor microenvironment [UMCN 1.3]
Alternative Splicing
Microscopy
Fluorescence
Biochemistry
COS Cells
Mutation
Electrophoresis
Polyacrylamide Gel
Plasmids
Zdroj: Molecular and Cellular Biology, 23, 16, pp. 5489-501
Molecular and Cellular Biology, 23, 5489-501
ISSN: 0270-7306
DOI: 10.1128/mcb.23.16.5489-5501.2003
Popis: Item does not contain fulltext Transcripts of the myotonic dystrophy protein kinase (DMPK) gene, a member of the Rho kinase family, are subject to cell-type specific alternative splicing. An imbalance in the splice isoform profile of DMPK may play a role in the pathogenesis of DM1, a severe multisystemic disorder. Here, we report how structural subdomains determine biochemical properties and subcellular distribution of DMPK isoforms. A newly developed kinase assay revealed that DMPK is a Lys/Arg-directed kinase. Individual DMPK isoforms displayed comparable transphosphorylation activity and sequence preference for peptide substrates. However, DMPK autophosphorylation and phosphorylation of MYPT1 (as putative in vivo target of DMPK), were dependent on presence of an alternatively spliced VSGGG motif and the nature of the C terminus. In-gel effects of the VSGGG motif on the migration behavior of full-length kinase provide evidence for a model in which this motif mediates 3-D-conformational changes in DMPK isoforms. Finally, different C termini endow DMPK with the ability to bind to either endoplasmic reticulum or mitochondria or to adopt a cytosolic location. Our results suggest that DMPK isoforms have cell-type and location dependent substrate specificities with a role in organellar and cytoarchitectural dynamics.
Databáze: OpenAIRE
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