The Mechanism of Tail-Anchored Protein Insertion into the ER Membrane

Fei Wang, Andrew Whynot, Matthew Tung, Vladimir Denic

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

Tail-anchored (TA) proteins access the secretory pathway via posttranslational insertion of their C-terminal transmembrane domain into the endoplasmic reticulum (ER). Get3 is an ATPase that delivers TA proteins to the ER by interacting with the Get1-Get2 transmembrane complex, but how Get3's nucleotide cycle drives TA protein insertion remains unclear. Here, we establish that nucleotide binding to Get3 promotes Get3-TA protein complex formation by recruiting Get3 to a chaperone that hands over TA proteins to Get3. Biochemical reconstitution and mutagenesis reveal that the Get1-Get2 complex comprises the minimal TA protein insertion machinery with functionally critical cytosolic regions. By engineering a soluble heterodimer of Get1-Get2 cytosolic domains, we uncover the mechanism of TA protein release from Get3: Get2 tethers Get3-TA protein complexes into proximity with the ATPase-dependent, substrate-releasing activity of Get1. Lastly, we show that ATP enhances Get3 dissociation from the membrane, thus freeing Get1-Get2 for new rounds of substrate insertion.

Original languageEnglish (US)
Pages (from-to)738-750
Number of pages13
JournalMolecular Cell
Volume43
Issue number5
DOIs
StatePublished - Sep 2 2011

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Endoplasmic Reticulum
Membranes
Proteins
Adenosine Triphosphatases
Nucleotides
Secretory Pathway
Mutagenesis
Hand
Adenosine Triphosphate

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

The Mechanism of Tail-Anchored Protein Insertion into the ER Membrane. / Wang, Fei; Whynot, Andrew; Tung, Matthew; Denic, Vladimir.

In: Molecular Cell, Vol. 43, No. 5, 02.09.2011, p. 738-750.

Research output: Contribution to journalArticle

Wang, Fei ; Whynot, Andrew ; Tung, Matthew ; Denic, Vladimir. / The Mechanism of Tail-Anchored Protein Insertion into the ER Membrane. In: Molecular Cell. 2011 ; Vol. 43, No. 5. pp. 738-750.
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