Lim C R, Kimata Y, Oka M, Nomaguchi K, Kohno K
Research and Education Center for Genetic Information, Nara Institute of Science and Technology.
J Biochem. 1995 Jul;118(1):13-7. doi: 10.1093/oxfordjournals.jbchem.a124868.
Tagging proteins with the green fluorescent protein (GFP) from Aequorea victoria is a good means of analyzing protein localization in living cells. Nevertheless, GFP and a chimeric protein, GFP-nucleoplasmin, expressed in Saccharomyces cerevisiae were less fluorescent at high culture temperatures. Proteins synthesized at a low temperature retained their fluorescence despite a shift to a higher temperature. Hence, when a temperature-sensitive nsp1 mutant expressing GFP-nucleoplasmin was cultured at 23 degrees C and then shifted to 35 degrees C, we were able to exclusively monitor the localization of the protein synthesized prior to the temperature shift. This protein accumulated in novel nuclear-like compartments devoid of DNA.
