Skip to main content
Dryad logo

Dataset associated with: The ERM-1 membrane-binding domain directs erm-1 mRNA localization to the plasma membrane in the C. elegans embryo

Citation

Winkenbach, Lindsay; Parker, Dylan; Williams, Robert; Osborne Nishimura, Erin (2022), Dataset associated with: The ERM-1 membrane-binding domain directs erm-1 mRNA localization to the plasma membrane in the C. elegans embryo, Dryad, Dataset, https://doi.org/10.5061/dryad.9cnp5hqnp

Abstract

mRNA localization and transport are integral in regulating gene expression. In Caenorhabditis elegans' embryos, the maternally inherited mRNA erm-1 (Ezrin/Radixin/Moesin) becomes concentrated in anterior blastomeres. erm-1 mRNA localizes within those blastomeres to the plasma membrane where the essential ERM-1 protein, a membrane-actin linker, is also found. We demonstrate that the localization of erm-1 mRNA to the plasma membrane is translation dependent and requires its encoded N-terminal, membrane-binding (FERM) domain. By perturbing translation through multiple methods, we found that erm-1 mRNA localization at the plasma membrane persisted only if the nascent peptide remained in complex with the translating mRNA. Indeed, re-coding the erm-1 mRNA coding sequence while preserving the encoded amino acid sequence did not disrupt erm-1 mRNA localization, corroborating that the information directing mRNA localization resides within its membrane-binding protein domain. A single-molecule inexpensive fluorescence in situ hybridization screen of 17 genes encoding similar membrane-binding domains identified three plasma membrane-localized mRNAs in the early embryo. Ten additional transcripts showed potential membrane localization later in development. These findings point to a translation-dependent pathway for localization of mRNAs encoding membrane-associated proteins.

Methods

All images were acquired on a Photometrics Cool Snap HQ2 camera using a DeltaVision Elite inverted microscope (GE Healthcare), with an Olympus PLAN APO 60X (1.42 NA, PLAPON60XOSC2) objective, an Insight SSI 7-Color Solid State Light Engine, and SoftWorx software (Applied Precision) using 0.2 um z-stacks.

Usage Notes

This dataset contains all primary microscopy data presented or analyzed in the publication "The ERM-1 membrane-binding domain directs erm-1 mRNA localization to the plasma membrane in the C. elegans embryo".

Funding

National Institutes of Health, Award: R25GM124877

National Science Foundation, Award: MCB CAREER award 2143849

National Science Foundation, Award: DGE-1450032