Characterization of photolyase/blue-light receptor homologs in mouse and human cells.
We isolated and characterized mouse photolyase-like genes, mCRY1 (mPHLL1) and mCRY2 (mPHLL2), which belong to the photolyase family including plant blue-light receptors. The mCRY1 and mCRY2 genes are located on chromosome 10C and 2E, respectively, and are expressed in all mouse organs examined. We raised antibodies specific against each gene product using its C-terminal sequence, which differs completely between the genes. Immunofluorescent staining of cultured mouse cells revealed that mCRY1 is localized in mitochondria whereas mCRY2 was found mainly in the nucleus. The subcellular distribution of CRY proteins was confirmed by immunoblot analysis of fractionated mouse liver cell extracts. Using green fluorescent protein fused peptides we showed that the C-terminal region of the mouse CRY2 protein contains a unique nuclear localization signal, which is absent in the CRY1 protein. The N-terminal region of CRY1 was shown to contain the mitochondrial transport signal. Recombinant as well as native CRY1 proteins from mouse and human cells showed a tight binding activity to DNA Sepharose, while CRY2 protein did not bind to DNA Sepharose at all under the same condition as CRY1. The different cellular localization and DNA binding properties of the mammalian photolyase homologs suggest that despite the similarity in the sequence the two proteins have distinct function(s).
Kobayashi
K
K
Department of Molecular Genetics, Institute of Development, Aging and Cancer, Tohoku University, 980 8575 Sendai, Japan.
Kanno
S
S
Smit
B
B
van der Horst
G T
GT
Takao
M
M
Yasui
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001483:Base Sequence; D002874:Chromosome Mapping; D056931:Cryptochromes; D004247:DNA / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D004255:Deoxyribodipyrimidine Photo-Lyase / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D029721:Drosophila Proteins; D005136:Eye Proteins; D005420:Flavoproteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals / Q000235:genetics; D017956:Photoreceptor Cells, Invertebrate; D011485:Protein Binding; D011817:Rabbits; D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D000906:Antibodies; C538963:CRY1 protein, human; C538966:CRY2 protein, human; C538964:Cry1 protein, mouse; C538967:Cry2 protein, mouse; D056931:Cryptochromes; D017931:DNA Primers; D029721:Drosophila Proteins; D005136:Eye Proteins; D005420:Flavoproteins; D019913:Nuclear Localization Signals; D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins; C119828:cry protein, Drosophila; D004247:DNA; D004255:Deoxyribodipyrimidine Photo-Lyase