PUBLICATION
Functional and structural analyses of cryptochrome: Vertebrate CRY regions responsible for interaction with the CLOCK: BMAL1 heterodimer and its nuclear localization
- Authors
- Hirayama, J., Nakamura, H., Ishikawa, T., Kobayashi, Y., and Todo, T.
- ID
- ZDB-PUB-030728-24
- Date
- 2003
- Source
- The Journal of biological chemistry 278(37): 35630-35628 (Journal)
- Registered Authors
- Hirayama, Jun
- Keywords
- none
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Binding Sites
- Biological Clocks
- Cryptochromes
- DNA Primers
- Drosophila Proteins*
- Escherichia coli/metabolism
- Eye Proteins*
- Flavoproteins/chemistry*
- Flavoproteins/genetics*
- Flavoproteins/metabolism
- Mice
- Models, Molecular
- Molecular Sequence Data
- Photoreceptor Cells, Invertebrate*
- Protein Conformation
- Receptors, G-Protein-Coupled
- Recombinant Fusion Proteins/chemistry
- Recombinant Fusion Proteins/metabolism
- Repressor Proteins/metabolism
- Restriction Mapping
- Sequence Alignment
- Sequence Homology, Amino Acid
- Transcription, Genetic
- Zebrafish
- PubMed
- 12832412 Full text @ J. Biol. Chem.
Citation
Hirayama, J., Nakamura, H., Ishikawa, T., Kobayashi, Y., and Todo, T. (2003) Functional and structural analyses of cryptochrome: Vertebrate CRY regions responsible for interaction with the CLOCK: BMAL1 heterodimer and its nuclear localization. The Journal of biological chemistry. 278(37):35630-35628.
Abstract
Mouse mCRY1 and zebrafish zCRY1a and zCRY3 belong to the DNA photolyase/Cryptochrome family. mCRY1 and zCRY1a repress CLOCK:BMAL1- mediated transcription, whereas zCRY3 does not. Reciprocal chimeras between zCRY1a and zCRY3 were generated to determine the zCRY1a regions responsible for nuclear translocation, interaction with the CLOCK:BMAL1 heterodimer, and repression of CLOCK:BMAL1- mediated transcription. Three regions, RD-2a (126-196), RD-1 (197-263), and RD-2b (264-293), were identified. Proteins in this family consist of an N-terminal a/b domain and a C-terminal helical domain connected by an interdomain loop. RD-2a is within this loop, RD-1 is at the N-terminal 50, and RD-2b at the following 31 amino acid residues of the helical domain. Either RD-2a or RD-1 is required for interaction with the CLOCK:BMAL1 heterodimer, and either RD-1 or RD-2b is required for the nuclear translocation of CRY. Both of these functions are prerequisites for the transcriptional repressor activity. The functional nuclear localizing signal (NLS) in the RD-2b region also was identified. The sequence is well-conserved among repressor-type CRYs, including mCRY1. Mutations in the NLS of mCRY1 reduce the extent of its nuclear localization. These findings show that both nuclear localization and interaction with the CLOCK:BMAL heterodimer are essential for transcriptional repression by CRY.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping