Nuclear localization of the Arabidopsis blue light receptor cryptochrome 2.
The cryptochrome blue light photoreceptor family of Arabidopsis thaliana consists of two members, CRY1 and CRY2 (PHH1). CRY2 contains a putative nuclear localization signal (NLS) within its C-terminal region. We examined whether CRY2 is localized in the nucleus and whether the C-terminal region of CRY2 is involved in nuclear targeting. Total cellular and nuclear protein extracts from Arabidopsis were subjected to immunoblot analysis with CRY2-specific antibodies. Strong CRY2 signals were obtained in the nuclear fraction. Fusion proteins consisting of the green fluorescent protein (GFP) and different fragments of CRY2 were expressed in parsley protoplasts and the localization of the fusion proteins was determined by fluorescence and confocal laser scanning microscopy. GFP-fusions containing the entire CRY2 protein or its C-terminal region were found exclusively in the nucleus. We conclude from these results that CRY2 is localized in the nucleus and that nuclear localization is mediated by the C-terminal region of CRY2.
Kleiner
O
O
FB Biologie/Botanik, Philipps-Universität, Marburg, Germany.
Kircher
S
S
Harter
K
K
Batschauer
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects*; D029681:Arabidopsis Proteins; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D056931:Cryptochromes; D017931:DNA Primers / Q000235:genetics; D029721:Drosophila Proteins; D005136:Eye Proteins; D005420:Flavoproteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008027:Light; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals; D017956:Photoreceptor Cells, Invertebrate; D045322:Photosynthetic Reaction Center Complex Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D030821:Plants, Genetically Modified; D011817:Rabbits; D043562:Receptors, G-Protein-Coupled; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D029681:Arabidopsis Proteins; C538962:CRY1 protein, Arabidopsis; D056931:Cryptochromes; D017931:DNA Primers; D029721:Drosophila Proteins; D005136:Eye Proteins; D005420:Flavoproteins; D019913:Nuclear Localization Signals; D045322:Photosynthetic Reaction Center Complex Proteins; D043562:Receptors, G-Protein-Coupled; D011993:Recombinant Fusion Proteins; C119828:cry protein, Drosophila