Nuclear Translation Visualized by Ribosome-bound Nascent Chain Puromycylation

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 2012 Apr 5

PMID 22472439

Citations 166

Authors

Alexandre David

Brian P Dolan

Heather D Hickman

Jonathan J Knowlton

Giovanna Clavarino

Philippe Pierre

Jack R Bennink

Jonathan W Yewdell

Affiliations

Soon will be listed here.

Abstract

Whether protein translation occurs in the nucleus is contentious. To address this question, we developed the ribopuromycylation method (RPM), which visualizes translation in cells via standard immunofluorescence microscopy. The RPM is based on ribosome-catalyzed puromycylation of nascent chains immobilized on ribosomes by antibiotic chain elongation inhibitors followed by detection of puromycylated ribosome-bound nascent chains with a puromycin (PMY)-specific monoclonal antibody in fixed and permeabilized cells. The RPM correlates localized translation with myriad processes in cells and can be applied to any cell whose translation is sensitive to PMY. In this paper, we use the RPM to provide evidence for translation in the nucleoplasm and nucleolus, which is regulated by infectious and chemical stress.

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