Switch to
Predicate | Object |
---|---|
rdf:type | |
lifeskim:mentions | |
pubmed:issue |
2
|
pubmed:dateCreated |
1992-4-20
|
pubmed:abstractText |
The three 21-bp repeats (Tax-responsive elements) of the long terminal repeat (LTR) of the human T-cell leukemia virus (HTLV-I) mediates the response of the Tax protein. All three Tax-responsive elements (TREs) contain a TGACG motif, reminiscent of the CREB/ATF-binding site TGACGTCA. DNA-affinity chromatography with the 5'-TRE resulted in a previous study in proteins of about 32, 36 to 42, 50 and 110 kDa. Here we demonstrate that the 42 kDa protein is the cAMP-response element-binding (CREB) protein. This is shown by phosphorylation of the proteins eluted from the DNA-affinity column with protein kinase A (PKA) in vitro and subsequent indirect immunoprecipitation with a CREB-specific antiserum raised against an internal CREB-specific peptide. This method allows detection of phosphorylated proteins by autoradiography with high sensitivity and is superior to metabolic labeling. One of the phosphorylated proteins co-migrates with immuno-affinity-purified CREB protein--also phosphorylated in vitro--and competes with the peptide antigen, which proves the specificity of the reaction. The purified CREB protein leads to specific DNA-protein complexes in DNA mobility-shift analyses with all three TREs. Comparison of these TRE-CREB complexes with those formed by nuclear extracts from the HTLV-I-transformed T-cell line C81-66-45 indicates that additional cellular factors contribute to the complexes, especially to the middle TRE. This is also shown by using CREB-depleted instead of complete nuclear extracts for DNA mobility-shift assays. Antibodies against CREB but not Tax affect the mobility of the DNA-protein complex.
|
pubmed:language |
eng
|
pubmed:journal | |
pubmed:citationSubset |
IM
|
pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Cyclic AMP Response...,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Viral,
http://linkedlifedata.com/resource/pubmed/chemical/DNA-Binding Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Nuclear Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Oligodeoxyribonucleotides
|
pubmed:status |
MEDLINE
|
pubmed:month |
Feb
|
pubmed:issn |
0950-9232
|
pubmed:author | |
pubmed:issnType |
Print
|
pubmed:volume |
7
|
pubmed:owner |
NLM
|
pubmed:authorsComplete |
Y
|
pubmed:pagination |
257-62
|
pubmed:dateRevised |
2006-11-15
|
pubmed:meshHeading |
pubmed-meshheading:1532242-Base Sequence,
pubmed-meshheading:1532242-Chromatography, Affinity,
pubmed-meshheading:1532242-Cyclic AMP Response Element-Binding Protein,
pubmed-meshheading:1532242-DNA, Viral,
pubmed-meshheading:1532242-DNA-Binding Proteins,
pubmed-meshheading:1532242-Gene Expression Regulation, Viral,
pubmed-meshheading:1532242-Human T-lymphotropic virus 1,
pubmed-meshheading:1532242-Humans,
pubmed-meshheading:1532242-Molecular Sequence Data,
pubmed-meshheading:1532242-Nuclear Proteins,
pubmed-meshheading:1532242-Oligodeoxyribonucleotides,
pubmed-meshheading:1532242-Regulatory Sequences, Nucleic Acid,
pubmed-meshheading:1532242-Repetitive Sequences, Nucleic Acid
|
pubmed:year |
1992
|
pubmed:articleTitle |
Direct interaction of CREB protein with 21 bp Tax-response elements of HTLV-ILTR.
|
pubmed:affiliation |
Max-Planck-Institut für Molekulare Genetik, Abt. Schuster, Berlin, Germany.
|
pubmed:publicationType |
Journal Article,
In Vitro,
Research Support, Non-U.S. Gov't
|