Hhat is a Palmitoylacyltransferase with Specificity for N-palmitoylation of Sonic Hedgehog

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2008 Jun 7

PMID 18534984

Citations 132

Authors

John A Buglino

Marilyn D Resh

Affiliations

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Abstract

Palmitoylation of Sonic Hedgehog (Shh) is critical for effective long- and short-range signaling. Genetic screens uncovered a potential palmitoylacyltransferase (PAT) for Shh, Hhat, but the molecular mechanism of Shh palmitoylation remains unclear. Here, we have developed and exploited an in vitro Shh palmitoylation assay to purify Hhat to homogeneity. We provide direct biochemical evidence that Hhat is a PAT with specificity for attaching palmitate via amide linkage to the N-terminal cysteine of Shh. Other palmitoylated proteins (e.g. PSD95 and Wnt) are not substrates for Hhat, and Porcupine, a putative Wnt PAT, does not palmitoylate Shh. Neither autocleavage nor cholesterol modification is required for Shh palmitoylation. Both the Shh precursor and mature protein are N-palmitoylated by Hhat, and the reaction occurs during passage through the secretory pathway. This study establishes Hhat as a bona fide Shh PAT and serves as a model for understanding how secreted morphogens are modified by distinct PATs.

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References

Hall T, Porter J, Beachy P, LEAHY D . A potential catalytic site revealed by the 1.7-A crystal structure of the amino-terminal signalling domain of Sonic hedgehog. Nature. 1995; 378(6553):212-6. DOI: 10.1038/378212a0. View

Chen M, Li Y, Kawakami T, Xu S, Chuang P . Palmitoylation is required for the production of a soluble multimeric Hedgehog protein complex and long-range signaling in vertebrates. Genes Dev. 2004; 18(6):641-59. PMC: 387240. DOI: 10.1101/gad.1185804. View

Gallet A, Rodriguez R, Ruel L, Therond P . Cholesterol modification of hedgehog is required for trafficking and movement, revealing an asymmetric cellular response to hedgehog. Dev Cell. 2003; 4(2):191-204. DOI: 10.1016/s1534-5807(03)00031-5. View

Kohtz J, Lee H, Gaiano N, Segal J, Ng E, Larson T . N-terminal fatty-acylation of sonic hedgehog enhances the induction of rodent ventral forebrain neurons. Development. 2001; 128(12):2351-63. DOI: 10.1242/dev.128.12.2351. View

Pepinsky R, Zeng C, Wen D, Rayhorn P, Baker D, Williams K . Identification of a palmitic acid-modified form of human Sonic hedgehog. J Biol Chem. 1998; 273(22):14037-45. DOI: 10.1074/jbc.273.22.14037. View

Thayer S, Pasca di Magliano M, Heiser P, Nielsen C, Roberts D, Lauwers G . Hedgehog is an early and late mediator of pancreatic cancer tumorigenesis. Nature. 2003; 425(6960):851-6. PMC: 3688051. DOI: 10.1038/nature02009. View

Llopis J, McCaffery J, Miyawaki A, Farquhar M, Tsien R . Measurement of cytosolic, mitochondrial, and Golgi pH in single living cells with green fluorescent proteins. Proc Natl Acad Sci U S A. 1998; 95(12):6803-8. PMC: 22642. DOI: 10.1073/pnas.95.12.6803. View

Callejo A, Torroja C, Quijada L, Guerrero I . Hedgehog lipid modifications are required for Hedgehog stabilization in the extracellular matrix. Development. 2006; 133(3):471-83. DOI: 10.1242/dev.02217. View

Gallet A, Ruel L, Staccini-Lavenant L, Therond P . Cholesterol modification is necessary for controlled planar long-range activity of Hedgehog in Drosophila epithelia. Development. 2006; 133(3):407-18. DOI: 10.1242/dev.02212. View

10.

Kleuss C, Krause E . Galpha(s) is palmitoylated at the N-terminal glycine. EMBO J. 2003; 22(4):826-32. PMC: 145455. DOI: 10.1093/emboj/cdg095. View

11.

Berthiaume L, Peseckis S, Resh M . Synthesis and use of iodo-fatty acid analogs. Methods Enzymol. 1995; 250:454-66. DOI: 10.1016/0076-6879(95)50090-1. View

12.

Schjerling C, Hummel R, Hansen J, Borsting C, Mikkelsen J, Kristiansen K . Disruption of the gene encoding the acyl-CoA-binding protein (ACB1) perturbs acyl-CoA metabolism in Saccharomyces cerevisiae. J Biol Chem. 1996; 271(37):22514-21. DOI: 10.1074/jbc.271.37.22514. View

13.

Mitchell D, Vasudevan A, Linder M, Deschenes R . Protein palmitoylation by a family of DHHC protein S-acyltransferases. J Lipid Res. 2006; 47(6):1118-27. DOI: 10.1194/jlr.R600007-JLR200. View

14.

Porter J, Young K, Beachy P . Cholesterol modification of hedgehog signaling proteins in animal development. Science. 1996; 274(5285):255-9. DOI: 10.1126/science.274.5285.255. View

15.

Chamoun Z, MANN R, Nellen D, von Kessler D, Bellotto M, Beachy P . Skinny hedgehog, an acyltransferase required for palmitoylation and activity of the hedgehog signal. Science. 2001; 293(5537):2080-4. DOI: 10.1126/science.1064437. View

16.

Farazi T, Waksman G, Gordon J . The biology and enzymology of protein N-myristoylation. J Biol Chem. 2001; 276(43):39501-4. DOI: 10.1074/jbc.R100042200. View

17.

Roth A, Wan J, Bailey A, Sun B, Kuchar J, Green W . Global analysis of protein palmitoylation in yeast. Cell. 2006; 125(5):1003-13. PMC: 2246083. DOI: 10.1016/j.cell.2006.03.042. View

18.

Lee J, Kraus P, Gaiano N, Nery S, Kohtz J, Fishell G . An acylatable residue of Hedgehog is differentially required in Drosophila and mouse limb development. Dev Biol. 2001; 233(1):122-36. DOI: 10.1006/dbio.2001.0218. View

19.

Ho K, Scott M . Sonic hedgehog in the nervous system: functions, modifications and mechanisms. Curr Opin Neurobiol. 2002; 12(1):57-63. DOI: 10.1016/s0959-4388(02)00290-8. View

20.

Hofmann K . A superfamily of membrane-bound O-acyltransferases with implications for wnt signaling. Trends Biochem Sci. 2000; 25(3):111-2. DOI: 10.1016/s0968-0004(99)01539-x. View