2021
DOI: 10.1371/journal.pbio.3001191
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Smoothened transduces Hedgehog signals via activity-dependent sequestration of PKA catalytic subunits

Abstract: The Hedgehog (Hh) pathway is essential for organ development, homeostasis, and regeneration. Dysfunction of this cascade drives several cancers. To control expression of pathway target genes, the G protein–coupled receptor (GPCR) Smoothened (SMO) activates glioma-associated (GLI) transcription factors via an unknown mechanism. Here, we show that, rather than conforming to traditional GPCR signaling paradigms, SMO activates GLI by binding and sequestering protein kinase A (PKA) catalytic subunits at the membran… Show more

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Cited by 47 publications
(124 citation statements)
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“…Alexa 647-conjugated M1 FLAG antibody and M1 FLAG affinity resin were prepared inhouse. Control or ShhN conditioned medium was prepared from stably transfected HEK293 cells as previously described 24 . Dual-luciferase assay was obtained from Promega.…”
Section: Methodsmentioning
confidence: 99%
See 4 more Smart Citations
“…Alexa 647-conjugated M1 FLAG antibody and M1 FLAG affinity resin were prepared inhouse. Control or ShhN conditioned medium was prepared from stably transfected HEK293 cells as previously described 24 . Dual-luciferase assay was obtained from Promega.…”
Section: Methodsmentioning
confidence: 99%
“…Proteins were separated by SDS-PAGE on a 4-20% Stain-Free TGX gel (BioRad), and total protein in lysate or eluate was visualized via Stain Free imaging. To detect PKA-Ca-YFP, inputs and eluates were transferred to PVDF membranes and processed for immunoblotting with anti-GFP antibodies as previously described 24 . In-gel Pro-Q Diamond assay to detect phosphoprotein was performed according to the manufacturer's instructions, as previously described 24 .…”
Section: Methodsmentioning
confidence: 99%
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