a systematic approach to pair secretory cargo receptors with their cargo suggests a mechanism for cargo selection by erv14系统方法对分泌货物受体与货物显示货物由erv14选择机制.pdfVIP

A Systematic Approach to Pair Secretory Cargo Receptors with Their Cargo Suggests a Mechanism for Cargo Selection by Erv14 1. 2. 1 2 1 Yonatan Herzig , Hayley J. Sharpe , Yael Elbaz , Sean Munro *, Maya Schuldiner * 1 Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel, 2 MRC Laboratory of Molecular Biology, Cambridge, United Kingdom Abstract The endoplasmic reticulum (ER) is the site of synthesis of secreted and membrane proteins. To exit the ER, proteins are packaged into COPII vesicles through direct interaction with the COPII coat or aided by specific cargo receptors. Despite the fundamental role of such cargo receptors in protein traffic, only a few have been identified; their cargo spectrum is unknown and the signals they recognize remain poorly understood. We present here an approach we term ‘‘PAIRS’’ (pairing analysis of cargo receptors), which combines systematic genetic manipulations of yeast with automated microscopy screening, to map the spectrum of cargo for a known receptor or to uncover a novel receptor for a particular cargo. Using PAIRS we followed the fate of ,150 cargos on the background of mutations in nine putative cargo receptors and identified novel cargo for most of these receptors. Deletion of the Erv14 cargo receptor affected the widest range of cargo. Erv14 substrates have a wide array of functions and structures; however, they are all membrane-spanning proteins of the late secretory pathway or plasma membrane. Proteins residing in these organelles have longer transmembrane domains (TMDs). Detailed examination of one cargo supported the hypothesis that Erv14 dependency reflects the length rather than the sequence of the TMD. Th