A Conserved Di-Basic Motif of Drosophila Crumbs Contributes to Efficient ER Export

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Alexandra KumichelElisabeth Knust

Abstract

The Drosophila type I transmembrane protein Crumbs is an apical determinant required for the maintenance of apico-basal epithelial cell polarity. The level of Crumbs at the plasma membrane is crucial, but how it is regulated is poorly understood. In a genetic screen for regulators of Crumbs protein trafficking we identified Sar1, the core component of the coat protein complex II transport vesicles. sar1 mutant embryos show a reduced plasma membrane localization of Crumbs, a defect similar to that observed in haunted and ghost mutant embryos, which lack Sec23 and Sec24CD, respectively. By pulse-chase assays in Drosophila Schneider cells and analysis of protein transport kinetics based on Endoglycosidase H resistance we identified an RNKR motif in Crumbs, which contributes to efficient ER export. The motif identified fits the highly conserved di-basic RxKR motif and mediates interaction with Sar1. The RNKR motif is also required for plasma membrane delivery of transgene-encoded Crumbs in epithelial cells of Drosophila embryos. Our data are the first to show that a di-basic motif acts as a signal for ER exit of a type I plasma membrane protein in a metazoan organism.

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Citations

Jun 10, 2016·Traffic·Alvaro Román-Fernández, David M Bryant
Apr 15, 2017·Small GTPases·Francisco F Rodrigues, Tony J C Harris
Jan 10, 2016·Disease Models & Mechanisms·Arend W OvereemSven C D van IJzendoorn
Mar 11, 2017·The Journal of Cell Biology·Min LiuJosé Carlos Pastor-Pareja
Sep 18, 2021·Traffic·Sharanya ChatterjeeGad Frankel

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Methods Mentioned

BETA
GTPase
nucleotide exchange
Confocal microscopy
electrophoresis
phosphotransferase
transgenic
PCR
transfection

Software Mentioned

ZEN
Excel
ClustalW
Adobe Illustrator
Fiji
ImageQuant TL

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