Optogenetic control of nuclear protein export.

Dominik Niopek,Julia Roensch,Pierre Wehler,Barbara Di Ventura,Roland Eils

doi:10.1038/ncomms10624

Dominik Niopek, Julia Roensch + Show 3 more

Open Access

https://doi.org/10.1038/ncomms10624

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Abstract

Active nucleocytoplasmic transport is a key mechanism underlying protein regulation in eukaryotes. While nuclear protein import can be controlled in space and time with a portfolio of optogenetic tools, protein export has not been tackled so far. Here we present a light-inducible nuclear export system (LEXY) based on a single, genetically encoded tag, which enables precise spatiotemporal control over the export of tagged proteins. A constitutively nuclear, chromatin-anchored LEXY variant expands the method towards light inhibition of endogenous protein export by sequestering cellular CRM1 receptors. We showcase the utility of LEXY for cell biology applications by regulating a synthetic repressor as well as human p53 transcriptional activity with light. LEXY is a powerful addition to the optogenetic toolbox, allowing various novel applications in synthetic and cell biology.

Highlights

Active nucleocytoplasmic transport is a key mechanism underlying protein regulation in eukaryotes
Optogenetic tools that enable controlling with light the nuclear import of tagged proteins in mammalian cells and yeast have been reported[2,3,4,5,6], but no optogenetic tools are yet available to directly control protein export
light-inducible nuclear export system (LEXY) consists of an engineered LOV2 domain from Avena sativa phototropin-1 (AsLOV2), in which the C-terminal Ja helix was converted into an artificial NES

Summary

Introduction

Active nucleocytoplasmic transport is a key mechanism underlying protein regulation in eukaryotes. Optogenetic tools that enable controlling with light the nuclear import of tagged proteins in mammalian cells and yeast have been reported[2,3,4,5,6], but no optogenetic tools are yet available to directly control protein export Such a tool would have enormous application potential, for example, for regulating the activity of nuclear or cytoplasmic signalling molecules, and would complement the existing optogenetic toolset for control of nuclear import[2,3,4,5,6], protein dimerization[7] and oligomerization[8,9], membrane recruitment[10] and organelle transport and positioning[11]. To demonstrate the utility of LEXY for applications in synthetic and cell biology, we regulate synthetic repressors as well as the transcriptional activity of human p53 with light

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