3 years ago

Tail-Anchored Protein Insertion by a Single Get1/2 Heterodimer

Tail-Anchored Protein Insertion by a Single Get1/2 Heterodimer
Ronald S. Rock, Benjamin E. Zalisko, Robert J. Keenan, Vladimir Denic, Charlene Chan

Summary

The Get1/2 transmembrane complex drives the insertion of tail-anchored (TA) proteins from the cytosolic chaperone Get3 into the endoplasmic reticulum membrane. Mechanistic insight into how Get1/2 coordinates this process is confounded by a lack of understanding of the basic architecture of the complex. Here, we define the oligomeric state of full-length Get1/2 in reconstituted lipid bilayers by combining single-molecule and bulk fluorescence measurements with quantitative in vitro insertion analysis. We show that a single Get1/2 heterodimer is sufficient for insertion and demonstrate that the conserved cytosolic regions of Get1 and Get2 bind asymmetrically to opposing subunits of the Get3 homodimer. Altogether, our results define a simplified model for how Get1/2 and Get3 coordinate TA protein insertion.

Publisher URL: http://www.cell.com/cell-reports/fulltext/S2211-1247(17)31142-7

DOI: 10.1016/j.celrep.2017.08.035

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