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. 2017 Feb;18(2):110-122.
doi: 10.1111/tra.12459. Epub 2017 Jan 3.

Cargo selectivity of yeast sorting nexins

Björn D M Bean  1 , Michael Davey  1 , Elizabeth Conibear  1
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Free article

Cargo selectivity of yeast sorting nexins

Björn D M Bean et al. Traffic. 2017 Feb.
Free article

Abstract

Sorting nexins are PX domain-containing proteins that bind phospholipids and often act in membrane trafficking where they help to select cargo. However, the functions and cargo specificities of many sorting nexins are unknown. Here, a high-throughput imaging screen was used to identify new sorting nexin cargo in the yeast Saccharomyces cerevisiae. Deletions of 9 different sorting nexins were screened for mislocalization of a set of green fluorescent protein (GFP)-tagged membrane proteins found at the plasma membrane, Golgi or endosomes. This identified 27 proteins that require 1 or more sorting nexins for their correct localization, 23 of which represent novel sorting nexin cargo. Nine hits whose sorting was dependent on Snx4, the sorting nexin-containing retromer complex, or both retromer and Snx3, were examined in detail to search for potential sorting motifs. We identified cytosolic domains of Ear1, Ymd8 and Ymr010w that conferred retromer-dependent sorting on a chimeric reporter and identified conserved residues required for this sorting in a functional assay. This work defined a consensus sequence for retromer and Snx3-dependent sorting.

Keywords: Ear1; Endosome; Golgi; High-Throughput Microscopy; Retrograde Trafficking; Retromer; Snx3; Snx4; Sorting Nexin; Yeast; Ymd8; Ymr010w; Ymr253c.

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