Thermosensitivity of green fluorescent protein fluorescence utilized to reveal novel nuclear-like compartments in a mutant nucleoporin NSP1.
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.
Research and Education Center for Genetic Information, Nara Institute of Science and Technology.
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Lim
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Kimata
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Oka
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Nomaguchi
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Kohno
D000595:Amino Acid Sequence; D001483:Base Sequence; D002135:Calcium-Binding Proteins; D002451:Cell Compartmentation; D002467:Cell Nucleus; D005656:Fungal Proteins / Q000032:analysis*; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D009154:Mutation; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins / Q000032:analysis*; D012441:Saccharomyces cerevisiae / Q000737:chemistry; D029701:Saccharomyces cerevisiae Proteins; D013050:Spectrometry, Fluorescence; D013696:Temperature
D002135:Calcium-Binding Proteins; D005656:Fungal Proteins; D008164:Luminescent Proteins; C062748:NSP1 protein, S cerevisiae; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins