The interaction of nucleoside diphosphate kinase B with Gbetagamma dimers controls heterotrimeric G protein function

Hippe, H.J., Wolf, N.M., Abu-Taha, I., Mehringer, R., Just, S., Lutz, S., Niroomand, F., Postel, E.H., Katus, H.A., Rottbauer, W., and Wieland, T.
Proceedings of the National Academy of Sciences of the United States of America   106(38): 16269-16274 (Journal)
Registered Authors
Just, Steffen, Rottbauer, Wolfgang, Wolf, Nadine
cAMP, cardiac contractility, G proteins, NDPK, zebrafish
MeSH Terms
  • Amino Acid Sequence
  • Animals
  • Cells, Cultured
  • Cyclic AMP/metabolism
  • Embryo, Nonmammalian/embryology
  • Embryo, Nonmammalian/metabolism
  • Fibroblasts/cytology
  • Fibroblasts/metabolism
  • GTP-Binding Protein beta Subunits/chemistry
  • GTP-Binding Protein beta Subunits/genetics
  • GTP-Binding Protein beta Subunits/metabolism*
  • GTP-Binding Protein gamma Subunits/chemistry
  • GTP-Binding Protein gamma Subunits/genetics
  • GTP-Binding Protein gamma Subunits/metabolism*
  • Gene Expression Regulation, Developmental
  • Gene Knockdown Techniques
  • Heterotrimeric GTP-Binding Proteins/genetics
  • Heterotrimeric GTP-Binding Proteins/metabolism*
  • Heterotrimeric GTP-Binding Proteins/physiology
  • Humans
  • Immunoblotting
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Molecular Sequence Data
  • Myocardial Contraction/genetics
  • Myocardial Contraction/physiology
  • Myocardium/metabolism
  • NM23 Nucleoside Diphosphate Kinases/genetics
  • NM23 Nucleoside Diphosphate Kinases/metabolism*
  • Protein Binding
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Homology, Amino Acid
  • Zebrafish/embryology
  • Zebrafish/genetics
  • Zebrafish/metabolism
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism*
19805292 Full text @ Proc. Natl. Acad. Sci. USA
Heterotrimeric G proteins in physiological and pathological processes have been extensively studied so far. However, little is known about mechanisms regulating the cellular content and compartmentalization of G proteins. Here, we show that the association of nucleoside diphosphate kinase B (NDPK B) with the G protein betagamma dimer (Gbetagamma) is required for G protein function in vivo. In zebrafish embryos, morpholino-mediated knockdown of zebrafish NDPK B, but not NDPK A, results in a severe decrease in cardiac contractility. The depletion of NDPK B is associated with a drastic reduction in Gbeta(1)gamma(2) dimer expression. Moreover, the protein levels of the adenylyl cyclase (AC)-regulating Galpha(s) and Galpha(i) subunits as well as the caveolae scaffold proteins caveolin-1 and -3 are strongly reduced. In addition, the knockdown of the zebrafish Gbeta(1) orthologs, Gbeta(1) and Gbeta(1like), causes a cardiac phenotype very similar to that of NDPK B morphants. The loss of Gbeta(1)/Gbeta(1like) is associated with a down-regulation in caveolins, AC-regulating Galpha-subunits, and most important, NDPK B. A comparison of embryonic fibroblasts from wild-type and NDPK A/B knockout mice demonstrate a similar reduction of G protein, caveolin-1 and basal cAMP content in mammalian cells that can be rescued by re-expression of human NDPK B. Thus, our results suggest a role for the interaction of NDPK B with Gbetagamma dimers and caveolins in regulating membranous G protein content and maintaining normal G protein function in vivo.
Genes / Markers
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Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes