title: The structural basis of tail-anchored membrane protein recognition by Get3.
abstract: Targeting of newly synthesized membrane proteins to the endoplasmic reticulum is an essential cellular process. Most membrane proteins are recognized and targeted co-translationally by the signal recognition particle. However, nearly 5% of membrane proteins are 'tail-anchored' by a single carboxy-terminal transmembrane domain that cannot access the co-translational pathway. Instead, tail-anchored proteins are targeted post-translationally by a conserved ATPase termed Get3. The mechanistic basis for tail-anchored protein recognition or targeting by Get3 is not known. Here we present crystal structures of yeast Get3 in 'open' (nucleotide-free) and 'closed' (ADP.AlF(4)(-)-bound) dimer states. In the closed state, the dimer interface of Get3 contains an enormous hydrophobic groove implicated by mutational analyses in tail-anchored protein binding. In the open state, Get3 undergoes a striking rearrangement that disrupts the groove and shields its hydrophobic surfaces. These data provide a molecular mechanism for nucleotide-regulated binding and release of tail-anchored proteins during their membrane targeting by Get3.
authors.affiliation: Department of Biochemistry & Molecular Biology, The University of Chicago, Gordon Center for Integrative Science, Room W238, 929 East 57th Street, Chicago, Illinois 60637, USA.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Mateja
authors.forename: Anna
authors.initials: A
authors.lastname: Szlachcic
authors.forename: Maureen E
authors.initials: ME
authors.lastname: Downing
authors.forename: Malgorzata
authors.initials: M
authors.lastname: Dobosz
authors.forename: Malaiyalam
authors.initials: M
authors.lastname: Mariappan
authors.forename: Ramanujan S
authors.initials: RS
authors.lastname: Hegde
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Keenan
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D017607:Aluminum Compounds / Q000737:chemistry / Q000378:metabolism; D018360:Crystallography, X-Ray; D005459:Fluorides / Q000737:chemistry / Q000378:metabolism; D020662:Guanine Nucleotide Exchange Factors / Q000737:chemistry* / Q000378:metabolism*; D057927:Hydrophobic and Hydrophilic Interactions; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D017017:Methanococcus; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D055503:Protein Multimerization; D000069816:SEC Translocation Channels; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D017607:Aluminum Compounds; D020662:Guanine Nucleotide Exchange Factors; D008565:Membrane Proteins; D000069816:SEC Translocation Channels; D029701:Saccharomyces cerevisiae Proteins; C050992:tetrafluoroaluminate; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; C529048:Get3 protein, S cerevisiae; D005459:Fluorides
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