GONUTS has been updated to MW1.31 Most things seem to be working but be sure to report problems.

Have any questions? Please email us at ecoliwiki@gmail.com

PMID:20018846

From GONUTS
Jump to: navigation, search
Citation

Kotani, T, Iemura, S, Natsume, T, Kawakami, K and Yamashita, M (2010) Mys protein regulates protein kinase A activity by interacting with regulatory type Ialpha subunit during vertebrate development. J. Biol. Chem. 285:5106-16

Abstract

During embryonic development, protein kinase A (PKA) plays a key role in cell fate specification by antagonizing the Hedgehog (Hh) signaling pathway. However, the mechanism by which PKA activity is regulated remains unknown. Here we show that the Misty somites (Mys) protein regulates the level of PKA activity during embryonic development in zebrafish. We isolate PKA regulatory type Ialpha subunit (Prkar1a) as a protein interacting with Mys by pulldown assay in HEK293 cells followed by mass spectrometry analysis. We show an interaction between endogenous Mys and Prkar1a in the zebrafish embryo. Mys binds to Prkar1a in its C terminus region, termed PRB domain, and activates PKA in vitro. Conversely, knockdown of Mys in zebrafish embryos results in reduction in PKA activity. We also show that knockdown of Mys induces ectopic activation of Hh target genes in the eyes, neural tube, and somites downstream of Smoothened, a protein essential for transduction of Hh signaling activity. The altered patterning of gene expression is rescued by activation of PKA. Together, our results reveal a molecular mechanism of regulation of PKA activity that is dependent on a protein-protein interaction and demonstrate that PKA activity regulated by Mys is indispensable for negative regulation of the Hh signaling pathway in Hh-responsive cells.

Links

PubMed PMC2836113 Online version:10.1074/jbc.M109.070995

Keywords

Animals; Cell Line; Cyclic AMP-Dependent Protein Kinases/genetics; Cyclic AMP-Dependent Protein Kinases/metabolism; Embryo, Nonmammalian/metabolism; Forskolin/pharmacology; Humans; Immunoblotting; Immunoprecipitation; In Situ Hybridization; Mass Spectrometry; Oligonucleotides, Antisense/pharmacology; Protein Binding/drug effects; Veratrum Alkaloids/pharmacology; Zebrafish; Zebrafish Proteins/antagonists & inhibitors; Zebrafish Proteins/genetics; Zebrafish Proteins/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

DANRE:PRRC1

located_in

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DANRE:PRRC1

involved_in

GO:0034199: activation of protein kinase A activity

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DANRE:PRRC1

enables

GO:0034237: protein kinase A regulatory subunit binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

DANRE:PRRC1

located_in

GO:0005794: Golgi apparatus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

See Help:References for how to manage references in GONUTS.