Nuclear translocation and DNA recognition signals colocalized within the bZIP domain of cyclic adenosine 3',5'-monophosphate response element-binding protein CREB.

Details

Serval ID
serval:BIB_EF408E2EDB2B
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Nuclear translocation and DNA recognition signals colocalized within the bZIP domain of cyclic adenosine 3',5'-monophosphate response element-binding protein CREB.
Journal
Molecular Endocrinology
Author(s)
Waeber G., Habener J.F.
ISSN
0888-8809
Publication state
Published
Issued date
1991
Peer-reviewed
Oui
Volume
5
Number
10
Pages
1431-1438
Language
english
Notes
Publication types: Comparative Study ; Journal Article
Abstract
CREB is a cAMP-responsive nuclear DNA-binding protein that binds to cAMP response elements and stimulates gene transcription upon activation of the cAMP signalling pathway. The protein consists of an amino-terminal transcriptional transactivation domain and a carboxyl-terminal DNA-binding domain (bZIP domain) comprised of a basic region and a leucine zipper involved in DNA recognition and dimerization, respectively. Recently, we discovered a testis-specific transcript of CREB that contains an alternatively spliced exon encoding multiple stop codons. CREB encoded by this transcript is a truncated protein lacking the bZIP domain. We postulated that the antigen detected by CREB antiserum in the cytoplasm of germinal cells is the truncated CREB that must also lack its nuclear translocation signal (NTS). To test this hypothesis we prepared multiple expression plasmids encoding carboxyl-terminal deletions of CREB and transiently expressed them in COS-1 cells. By Western immunoblot analysis as well as immunocytochemistry of transfected cells, we show that CREB proteins truncated to amino acid 286 or shorter are sequestered in the cytoplasm, whereas a CREB of 295 amino acids is translocated into the nucleus. Chimeric CREBs containing a heterologous NTS fused to the first 248 or 261 amino acids of CREB are able to drive the translocation of the protein into the nucleus. Thus, the nine amino acids in the basic region involved in DNA recognition between positions 287 and 295 (RRKKKEYVK) of CREB contain the NTS. Further, mutation of the lysine at position 290 in CREB to an asparagine diminishes nuclear translocation of the protein.(ABSTRACT TRUNCATED AT 250 WORDS)
Keywords
Amino Acid Sequence, Animals, Base Sequence, Binding Sites, Cell Line, Cell Nucleus, Chromosome Deletion, Cyclic AMP Response Element-Binding Protein, DNA-Binding Proteins, Humans, Molecular Sequence Data, Mutagenesis, Site-Directed, Oligodeoxyribonucleotides, Plasmids, Polymerase Chain Reaction, RNA Splicing, Sequence Homology, Nucleic Acid, Transcription Factors, Transfection
Pubmed
Web of science
Open Access
Yes
Create date
25/01/2008 14:10
Last modification date
20/08/2019 16:17
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