PT - JOURNAL ARTICLE AU - Anatoly Meller AU - David Coombs AU - Reut Shalgi TI - Fluorescent polysome profiling reveals stress-mediated regulation of HSPA14-ribosome interactions AID - 10.1101/860833 DP - 2019 Jan 01 TA - bioRxiv PG - 860833 4099 - http://biorxiv.org/content/early/2019/12/01/860833.short 4100 - http://biorxiv.org/content/early/2019/12/01/860833.full AB - Ribosome-associated factors play important roles in regulation of translation in response to various physiological and environmental signals. Here we present fluorescent polysome profiling, a new method that provides simultaneous detection of UV and fluorescence directly from polysome gradients. We demonstrate the capabilities of the method by following the polysome incorporation of different fluorescently tagged ribosomal proteins in human cells. Additionally, we used fluorescent polysome profiling to examine chaperone-ribosome interactions, and characterized their changes in response to proteotoxic stresses. We revealed dynamic regulation of HSPA14-polysome association in response to heat shock, showing a marked heat shock-mediated increased in the polysome-association of HSPA14. Our data further support a model whereby HSPA14 dimerization is increased upon heat shock. We therefore established fluorescent polysome profiling as a powerful, streamlined method that can significantly enhance the study of ribosome-associated factors and their regulation.