title: A protein interaction network links GIT1, an enhancer of huntingtin aggregation, to Huntington's disease.
abstract: Analysis of protein-protein interactions (PPIs) is a valuable approach for characterizing proteins of unknown function. Here, we have developed a strategy combining library and matrix yeast two-hybrid screens to generate a highly connected PPI network for Huntington's disease (HD). The network contains 186 PPIs among 35 bait and 51 prey proteins. It revealed 165 new potential interactions, 32 of which were confirmed by independent binding experiments. The network also permitted the functional annotation of 16 uncharacterized proteins and facilitated the discovery of GIT1, a G protein-coupled receptor kinase-interacting protein, which enhances huntingtin aggregation by recruitment of the protein into membranous vesicles. Coimmunoprecipitations and immunofluorescence studies revealed that GIT1 and huntingtin associate in mammalian cells under physiological conditions. Moreover, GIT1 localizes to neuronal inclusions, and is selectively cleaved in HD brains, indicating that its distribution and function is altered during disease pathogenesis.
authors.affiliation: Max-Delbrueck-Center for Molecular Medicine, 13125 Berlin-Buch, Germany.
authors.forename: Heike
authors.initials: H
authors.lastname: Goehler
authors.forename: Maciej
authors.initials: M
authors.lastname: Lalowski
authors.forename: Ulrich
authors.initials: U
authors.lastname: Stelzl
authors.forename: Stephanie
authors.initials: S
authors.lastname: Waelter
authors.forename: Martin
authors.initials: M
authors.lastname: Stroedicke
authors.forename: Uwe
authors.initials: U
authors.lastname: Worm
authors.forename: Anja
authors.initials: A
authors.lastname: Droege
authors.forename: Katrin S
authors.initials: KS
authors.lastname: Lindenberg
authors.forename: Maria
authors.initials: M
authors.lastname: Knoblich
authors.forename: Christian
authors.initials: C
authors.lastname: Haenig
authors.forename: Martin
authors.initials: M
authors.lastname: Herbst
authors.forename: Jaana
authors.initials: J
authors.lastname: Suopanki
authors.forename: Eberhard
authors.initials: E
authors.lastname: Scherzinger
authors.forename: Claudia
authors.initials: C
authors.lastname: Abraham
authors.forename: Bianca
authors.initials: B
authors.lastname: Bauer
authors.forename: Renate
authors.initials: R
authors.lastname: Hasenbank
authors.forename: Anja
authors.initials: A
authors.lastname: Fritzsche
authors.forename: Andreas H
authors.initials: AH
authors.lastname: Ludewig
authors.forename: Konrad
authors.initials: K
authors.lastname: Büssow
authors.forename: Konrad
authors.initials: K
authors.lastname: Buessow
authors.forename: Sarah H
authors.initials: SH
authors.lastname: Coleman
authors.forename: Claire-Anne
authors.initials: CA
authors.lastname: Gutekunst
authors.forename: Bernhard G
authors.initials: BG
authors.lastname: Landwehrmeyer
authors.forename: Hans
authors.initials: H
authors.lastname: Lehrach
authors.forename: Erich E
authors.initials: EE
authors.lastname: Wanker
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001665:Binding Sites; D019556:COS Cells; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000378:metabolism*; D002522:Chlorocebus aethiops; D020690:GTPase-Activating Proteins / Q000737:chemistry / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D000071058:Huntingtin Protein; D006816:Huntington Disease / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008822:Mice, Transgenic; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D016716:PC12 Cells; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism*; D011233:Precipitin Tests; D011392:Proline / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014018:Tissue Distribution; D020798:Two-Hybrid System Techniques
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000911:Antibodies, Monoclonal; D018797:Cell Cycle Proteins; C485647:GIT1 protein, human; D020690:GTPase-Activating Proteins; C120766:Git1 protein, mouse; C485648:Git1 protein, rat; C086055:HTT protein, human; C506812:Htt protein, mouse; C499524:Htt protein, rat; D000071058:Huntingtin Protein; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; D011392:Proline; D005978:Glutathione