Distinct consequences of sterol sensor mutations in Drosophila and mouse patched homologs.

The membrane protein Patched (Ptc) is a critical regulator of Hedgehog signaling. Ptc is among a family of proteins that contain a sterol sensor motif. The function of this domain is poorly understood, but some proteins that contain sterol sensors are involved in cholesterol homeostasis. In the SREBP cleavage-activating protein (SCAP), sterols inhibit the protein's activity through this domain. Mutations in two highly conserved residues in the SCAP sterol sensor have been identified that confer resistance to sterol regulation. We introduced the analogous mutations in the sterol sensor motif of fly Ptc and mouse Ptc1 and examined their effect on protein activity. In contrast to SCAP, the sterol sensor mutations had different affects on Drosophila Ptc; Ptc Y442C retained function, while Ptc D584N conferred dominant negative activity. In the wing imaginal disc, Ptc D584N overexpression induced Hedgehog targets by stabilizing Cubitus interruptus and inducing decapentaplegic. However, Ptc D584N did not induce collier, a gene that requires high levels of Hedgehog signaling. In mouse Ptc1, the Y438C and D585N mutations did not stimulate signaling in Shh-responsive cell lines but did complement murine ptc1(-/-) cells. The results suggest that mutations in sterol sensor motifs alter function differently between sterol sensor family members.

[1]  Xin Zeng,et al.  A freely diffusible form of Sonic hedgehog mediates long-range signalling , 2001, Nature.

[2]  J. Taipale,et al.  The Hedgehog and Wnt signalling pathways in cancer , 2001, Nature.

[3]  G. Fishell,et al.  An acylatable residue of Hedgehog is differentially required in Drosophila and mouse limb development. , 2001, Developmental biology.

[4]  P. Ingham,et al.  Mutations in the Sterol Sensing Domain of Patched suggest a Role for Vesicular Trafficking in Smoothened Regulation , 2001, Current Biology.

[5]  Isabel Guerrero,et al.  The sterol-sensing domain of Patched protein seems to control Smoothened activity through Patched vesicular trafficking , 2001, Current Biology.

[6]  M. Scott,et al.  Effects of oncogenic mutations in Smoothened and Patched can be reversed by cyclopamine , 2000, Nature.

[7]  S. Cohen,et al.  Hedgehog Induces Opposite Changes in Turnover and Subcellular Localization of Patched and Smoothened , 2000, Cell.

[8]  R. L. Johnson,et al.  In vivo functions of the patched protein: requirement of the C terminus for target gene inactivation but not Hedgehog sequestration. , 2000, Molecular cell.

[9]  D. Robbins,et al.  Identification of a Tetrameric Hedgehog Signaling Complex* , 2000, The Journal of Biological Chemistry.

[10]  Michel Vervoort,et al.  hedgehog and wing development in Drosophila: a morphogen at work? , 2000, BioEssays : news and reviews in molecular, cellular and developmental biology.

[11]  C. Wicking,et al.  The hedgehog signalling pathway in tumorigenesis and development , 1999, Oncogene.

[12]  T. Kornberg,et al.  Ci: a complex transducer of the hedgehog signal. , 1999, Trends in genetics : TIG.

[13]  M. Brown,et al.  A proteolytic pathway that controls the cholesterol content of membranes, cells, and blood. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[14]  P. Beachy,et al.  Sonic hedgehog protein signals not as a hydrolytic enzyme but as an apparent ligand for patched. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[15]  H. Watari,et al.  Mutations in the Leucine Zipper Motif and Sterol-sensing Domain Inactivate the Niemann-Pick C1 Glycoprotein* , 1999, The Journal of Biological Chemistry.

[16]  Alain Vincent,et al.  The COE transcription factor Collier is a mediator of short-range Hedgehog-induced patterning of the Drosophila wing , 1999, Current Biology.

[17]  F. McCormick,et al.  A frequent activated smoothened mutation in sporadic basal cell carcinomas , 1999, Oncogene.

[18]  G. Struhl,et al.  In vivo evidence that Patched and Smoothened constitute distinct binding and transducing components of a Hedgehog receptor complex. , 1998, Development.

[19]  A. Rosenthal,et al.  Characterization of two patched receptors for the vertebrate hedgehog protein family. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[20]  H. Tricoire,et al.  Modulation of Hedgehog target gene expression by the Fused serine–threonine kinase in wing imaginal discs , 1998, Mechanisms of Development.

[21]  M. Brown,et al.  Sterols regulate processing of carbohydrate chains of wild-type SREBP cleavage-activating protein (SCAP), but not sterol-resistant mutants Y298C or D443N. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[22]  P. Ingham,et al.  Transducing Hedgehog: the story so far , 1998, The EMBO journal.

[23]  Q. Gu,et al.  Activating Smoothened mutations in sporadic basal-cell carcinoma , 1998, Nature.

[24]  M. Strigini,et al.  A Hedgehog activity gradient contributes to AP axial patterning of the Drosophila wing. , 1997, Development.

[25]  J. Mohler,et al.  Role of knot (kn) in wing patterning in Drosophila. , 1997, Genetics.

[26]  M. Scott,et al.  Altered neural cell fates and medulloblastoma in mouse patched mutants. , 1997, Science.

[27]  A. Yamaguchi,et al.  Induction of osteogenic differentiation by hedgehog proteins. , 1997, Biochemical and biophysical research communications.

[28]  M. Scott,et al.  Costal2, a Novel Kinesin-Related Protein in the Hedgehog Signaling Pathway , 1997, Cell.

[29]  R. Kobayashi,et al.  Hedgehog Elicits Signal Transduction by Means of a Large Complex Containing the Kinesin-Related Protein Costal2 , 1997, Cell.

[30]  W. Pavan,et al.  Murine model of Niemann-Pick C disease: mutation in a cholesterol homeostasis gene. , 1997, Science.

[31]  K. G. Coleman,et al.  Niemann-Pick C1 disease gene: homology to mediators of cholesterol homeostasis. , 1997, Science.

[32]  T. Kornberg,et al.  Proteolysis That Is Inhibited by Hedgehog Targets Cubitus interruptus Protein to the Nucleus and Converts It to a Repressor , 1997, Cell.

[33]  J. Mullor,et al.  Hedgehog activity, independent of decapentaplegic, participates in wing disc patterning. , 1997, Development.

[34]  M. Brown,et al.  Recurrent G-to-A substitution in a single codon of SREBP cleavage-activating protein causes sterol resistance in three mutant Chinese hamster ovary cell lines. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[35]  M. Scott,et al.  The tumour-suppressor gene patched encodes a candidate receptor for Sonic hedgehog , 1996, Nature.

[36]  C. Tabin,et al.  Biochemical evidence that Patched is the Hedgehog receptor , 1996, Nature.

[37]  X. Hua,et al.  Sterol Resistance in CHO Cells Traced to Point Mutation in SREBP Cleavage–Activating Protein , 1996, Cell.

[38]  G. Struhl,et al.  Dual Roles for Patched in Sequestering and Transducing Hedgehog , 1996, Cell.

[39]  Michael Dean,et al.  Is human patched the gatekeeper of common skin cancers? , 1996, Nature Genetics.

[40]  P. Ingham,et al.  Transcriptional activation of hedgehog target genes in Drosophila is mediated directly by the cubitus interruptus protein, a member of the GLI family of zinc finger DNA-binding proteins. , 1996, Genes & development.

[41]  P. Ingham,et al.  smoothened encodes a receptor-like serpentine protein required for hedgehog signalling , 1996, Nature.

[42]  M. Noll,et al.  The Drosophila smoothened Gene Encodes a Seven-Pass Membrane Protein, a Putative Receptor for the Hedgehog Signal , 1996, Cell.

[43]  Eugene V Koonin,et al.  Hedgehog Patterning Activity: Role of a Lipophilic Modification Mediated by the Carboxy-Terminal Autoprocessing Domain , 1996, Cell.

[44]  Michael Dean,et al.  Mutations of the Human Homolog of Drosophila patched in the Nevoid Basal Cell Carcinoma Syndrome , 1996, Cell.

[45]  R. Myers,et al.  Human Homolog of patched, a Candidate Gene for the Basal Cell Nevus Syndrome , 1996, Science.

[46]  M. Scott,et al.  Conservation of the hedgehog/patched signaling pathway from flies to mice: induction of a mouse patched gene by Hedgehog. , 1996, Genes & development.

[47]  R. L. Johnson,et al.  patched overexpression alters wing disc size and pattern: transcriptional and post-transcriptional effects on hedgehog targets. , 1995, Development.

[48]  E. Knust,et al.  Expression of crumbs confers apical character on plasma membrane domains of ectodermal epithelia of drosophila , 1995, Cell.

[49]  C. K. Motzny,et al.  The Drosophila cubitus interruptus protein and its role in the wingless and hedgehog signal transduction pathways , 1995, Mechanisms of Development.

[50]  T. Jessell,et al.  Floor plate and motor neuron induction by different concentrations of the amino-terminal cleavage product of sonic hedgehog autoproteolysis , 1995, Cell.

[51]  M. Scott,et al.  patched overexpression causes loss of wingless expression in Drosophila embryos. , 1994, Developmental biology.

[52]  Konrad Basler,et al.  Compartment boundaries and the control of Drosopfiffa limb pattern by hedgehog protein , 1994, Nature.

[53]  T. Hawley,et al.  Versatile retroviral vectors for potential use in gene therapy. , 1994, Gene therapy.

[54]  T. Tabata,et al.  Hedgehog is a signaling protein with a key role in patterning Drosophila imaginal discs , 1994, Cell.

[55]  E. Sánchez-Herrero,et al.  The Drosophila segment polarity gene patched interacts with decapentaplegic in wing development. , 1994, The EMBO journal.

[56]  N. Perrimon,et al.  Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. , 1993, Development.

[57]  P. Beachy,et al.  Secretion and localized transcription suggest a role in positional signaling for products of the segmentation gene hedgehog , 1992, Cell.

[58]  P. Ingham,et al.  Role of the Drosophila patched gene in positional signalling , 1991, Nature.

[59]  T. Préat,et al.  A putative serine/threonine protein kinase encoded by the segment-polarity fused gene of Drosophila , 1990, Nature.

[60]  K. Kroll,et al.  Cloning and characterization of the segment polarity gene cubitus interruptus Dominant of Drosophila. , 1990, Genes & development.

[61]  M. Scott,et al.  The Drosophila patched gene encodes a putative membrane protein required for segmental patterning , 1989, Cell.

[62]  A. Taylor,et al.  A protein with several possible membrane-spanning domains encoded by the Drosophila segment polarity gene patched , 1989, Nature.

[63]  Robert J. Gorlin,et al.  Nevoid Basal‐Cell Carcinoma Syndrome , 1987, Medicine.

[64]  B. Stevens,et al.  Human 3-hydroxy-3-methylglutaryl coenzyme A reductase. Conserved domains responsible for catalytic activity and sterol-regulated degradation. , 1985, The Journal of biological chemistry.

[65]  J. Goldstein,et al.  Membrane-bound domain of HMG CoA reductase is required for sterol-enhanced degradation of the enzyme , 1985, Cell.

[66]  G. Rubin,et al.  Transposition of cloned P elements into Drosophila germ line chromosomes. , 1982, Science.

[67]  E. Bier,et al.  Activation of knot (kn) specifies the 3-4 intervein region in the Drosophila wing. , 2000, Development.

[68]  Andrew P. McMahon,et al.  The world according to bedgebog , 1997 .

[69]  A. Felsenfeld,et al.  Positional signaling by hedgehog in Drosophila imaginal disc development. , 1995, Development.