RRC ID 38193
Author Asaoka Y, Kanai F, Ichimura T, Tateishi K, Tanaka Y, Ohta M, Seto M, Tada M, Ijichi H, Ikenoue T, Kawabe T, Isobe T, Yaffe MB, Omata M.
Title Identification of a suppressive mechanism for Hedgehog signaling through a novel interaction of Gli with 14-3-3.
Journal J Biol Chem
Abstract Gli transcription factors are central effectors of Hedgehog signaling in development and tumorigenesis. Using a tandem affinity purification (TAP) strategy and mass spectrometry, we have found that Gli1 interacts with 14-3-3epsilon, and that Gli2 and Gli3 also bind to 14-3-3epsilon through homologous sites. This interaction depends on their phosphorylation, and cAMP-dependent protein kinase (PKA), a known negative regulator of Hedgehog signaling serves as a responsible kinase. A Gli2 mutant engineered to eliminate this interaction exhibited increased transcriptional activity (2 approximately 3x). Transcriptional repression by 14-3-3 binding was also observed with Gli3, when its N-terminal repressor domain was deleted. The phosphorylation sites responsible for the binding to 14-3-3 are distinct from those required for proteolysis, the known mechanism for PKA-induced repression of Hh signaling. Our data propose a novel mechanism in which PKA down-regulates Hedgehog signaling by promoting the interaction between Gli and 14-3-3 as well as proteolysis. Given the certain neuronal or malignant disorders in human caused by the abnormality of 17p13 encompassing 14-3-3epsilon overlap with increased Hh signaling, the Gli-14-3-3 interaction may have pathological significance for those human diseases.
Volume 285(6)
Pages 4185-94
Published 2010-2-5
DOI 10.1074/jbc.M109.038232
PII M109.038232
PMID 19996099
PMC PMC2823557
MeSH 14-3-3 Proteins / genetics 14-3-3 Proteins / metabolism* Amino Acid Substitution Animals Binding Sites Cell Line Cell Line, Tumor Cyclic AMP-Dependent Protein Kinases / metabolism HeLa Cells Hedgehog Proteins / genetics Hedgehog Proteins / metabolism* Humans Immunoblotting Immunoprecipitation Kruppel-Like Transcription Factors / genetics Kruppel-Like Transcription Factors / metabolism Mass Spectrometry Mice NIH 3T3 Cells Nerve Tissue Proteins / genetics Nerve Tissue Proteins / metabolism Nuclear Proteins / genetics Nuclear Proteins / metabolism Phosphorylation Protein Binding RNA Interference Serine / genetics Serine / metabolism Signal Transduction* Transcription Factors / genetics Transcription Factors / metabolism* Transfection Zinc Finger Protein GLI1 Zinc Finger Protein Gli2 Zinc Finger Protein Gli3
IF 4.106
Times Cited 31
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Human and Animal Cells