Intraflagellar transport, cilia, and mammalian Hedgehog signaling: analysis in mouse embryonic fibroblasts.
Genetic studies in the mouse have shown that Intraflagellar Transport (IFT) is essential for mammalian Hedgehog (Hh) signal transduction. In this study, we take advantage of wild type and IFT mutant mouse embryonic fibroblasts (MEFs) to characterize additional aspects of the relationship between IFT and Hh signaling. Exposure to Sonic hedgehog (Shh) ligand or expression of an activated allele of Smo, SmoA1, activates an Hh reporter in wild-type MEFs, but not in MEFs derived from embryos that lack IFT172 or the Dync2h1 subunit of the retrograde IFT motor. Similarly, decreased activity of either Sufu or PKA, two negative regulators of Hh signal transduction, activates the pathway in wild-type, but not IFT mutant, MEFs. In contrast to wild-type MEFs, Smo is constitutively present in the cilia of Dync2h1 mutant MEFs. This finding suggests that IFT-dependent trafficking of Hh pathway components through the cilium is essential for their function.
Ocbina
Polloneal Jymmiel R
PJ
Developmental Biology Program, Sloan-Kettering Institute, New York, New York 10065, USA.
Anderson
Kathryn V
KV
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001692:Biological Transport / Q000502:physiology; D002923:Cilia / Q000378:metabolism* / Q000648:ultrastructure; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism; D003598:Cytoskeletal Proteins; D005333:Fetus / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism / Q000648:ultrastructure*; D005407:Flagella / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D053823:Hedgehog Proteins / Q000235:genetics* / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D008156:Luciferases / Q000235:genetics; D051379:Mice; D008817:Mice, Mutant Strains; D008855:Microscopy, Electron, Scanning; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D000072086:Smoothened Receptor
D048868:Adaptor Proteins, Signal Transducing; D003598:Cytoskeletal Proteins; D053823:Hedgehog Proteins; C479520:Ift172 protein, mouse; D047908:Intracellular Signaling Peptides and Proteins; D043562:Receptors, G-Protein-Coupled; D012097:Repressor Proteins; C465570:Shh protein, mouse; C491022:Smo protein, mouse; D000072086:Smoothened Receptor; C490763:Sufu protein, mouse; D008156:Luciferases; D017868:Cyclic AMP-Dependent Protein Kinases