title: Attenuation of hedgehog acyltransferase-catalyzed sonic Hedgehog palmitoylation causes reduced signaling, proliferation and invasiveness of human carcinoma cells.
abstract: Overexpression of Hedgehog family proteins contributes to the aetiology of many cancers. To be highly active, Hedgehog proteins must be palmitoylated at their N-terminus by the MBOAT family multispanning membrane enzyme Hedgehog acyltransferase (Hhat). In a pancreatic ductal adenocarcinoma (PDAC) cell line PANC-1 and transfected HEK293a cells Hhat localized to the endoplasmic reticulum. siRNA knockdown showed that Hhat is required for Sonic hedgehog (Shh) palmitoylation, for its assembly into high molecular weight extracellular complexes and for functional activity. Hhat knockdown inhibited Hh autocrine and juxtacrine signaling, and inhibited PDAC cell growth and invasiveness in vitro. In addition, Hhat knockdown in a HEK293a cell line constitutively expressing Shh and A549 human non-small cell lung cancer cells inhibited their ability to signal in a juxtacrine/paracrine fashion to the reporter cell lines C3H10T1/2 and Shh-Light2. Our data identify Hhat as a key player in Hh-dependent signaling and tumour cell transformed behaviour.
authors.affiliation: Molecular Medicine Section, National Heart & Lung Institute, Imperial College London, London, United Kingdom.
authors.forename: Antonios D
authors.initials: AD
authors.lastname: Konitsiotis
authors.affiliation: Molecular Medicine Section, National Heart & Lung Institute, Imperial College London, London, United Kingdom.
authors.forename: Shu-Chun
authors.initials: SC
authors.lastname: Chang
authors.affiliation: Molecular Medicine Section, National Heart & Lung Institute, Imperial College London, London, United Kingdom.
authors.forename: Biljana
authors.initials: B
authors.lastname: Jovanović
authors.affiliation: Department of Chemistry, Imperial College London, London, United Kingdom; Institute of Chemical Biology, Imperial College London, London, United Kingdom.
authors.forename: Paulina
authors.initials: P
authors.lastname: Ciepla
authors.affiliation: Department of Chemistry, Imperial College London, London, United Kingdom; Institute of Chemical Biology, Imperial College London, London, United Kingdom.
authors.forename: Naoko
authors.initials: N
authors.lastname: Masumoto
authors.affiliation: Institute for Health Research and Policy, London Metropolitan University, London, United Kingdom.
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Palmer
authors.affiliation: Department of Chemistry, Imperial College London, London, United Kingdom; Institute of Chemical Biology, Imperial College London, London, United Kingdom.
authors.forename: Edward W
authors.initials: EW
authors.lastname: Tate
authors.affiliation: Department of Biomedical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: John R
authors.initials: JR
authors.lastname: Couchman
authors.affiliation: Molecular Medicine Section, National Heart & Lung Institute, Imperial College London, London, United Kingdom; Institute of Chemical Biology, Imperial College London, London, United Kingdom.
authors.forename: Anthony I
authors.initials: AI
authors.lastname: Magee
mesh_terms: D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism*; D021441:Carcinoma, Pancreatic Ductal / Q000201:enzymology / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D053823:Hedgehog Proteins / Q000378:metabolism*; D006801:Humans; D054878:Lipoylation / Q000235:genetics / Q000502:physiology; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology
chemical_list: D053823:Hedgehog Proteins; D000217:Acyltransferases; C105599:HHAT protein, human