Publication Date:
1991-05-31
Description:
An in vivo selection system for isolating targets of DNA binding proteins in yeast was developed and used to identify the DNA binding site for the NGFI-B protein, a member of the steroid-thyroid hormone receptor superfamily. The feasibility of the technique was verified by selecting DNA fragments that contained binding sites for GCN4, a well-characterized yeast transcriptional activator. The DNA binding domain of NGFI-B, expressed as part of a LexA-NGFI-B-GAL4 chimeric activator, was then used to isolate a rat genomic DNA fragment that contained an NGFI-B binding site. The NGFI-B response element (NBRE) is similar to but functionally distinct from elements recognized by the estrogen and thyroid hormone receptors and the hormone receptor-like proteins COUP-TF, CF1, and H-2RIIBP. Cotransfection experiments in mammalian cells demonstrated that NGFI-B can activate transcription from the NBRE with or without its putative ligand binding domain.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Wilson, T E -- Fahrner, T J -- Johnston, M -- Milbrandt, J -- NS01018/NS/NINDS NIH HHS/ -- P01 CA49712/CA/NCI NIH HHS/ -- New York, N.Y. -- Science. 1991 May 31;252(5010):1296-300.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Pathology, Washington University School of Medicine, St. Louis, MO 63110.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/1925541" target="_blank"〉PubMed〈/a〉
Keywords:
Animals
;
Bacterial Proteins/metabolism
;
Base Sequence
;
Binding Sites
;
Cloning, Molecular
;
DNA, Fungal/*metabolism
;
DNA-Binding Proteins/genetics/*metabolism/pharmacology
;
Fungal Proteins/metabolism
;
Molecular Sequence Data
;
Nuclear Receptor Subfamily 4, Group A, Member 1
;
Plasmids
;
*Protein Kinases
;
Rats
;
Receptors, Cytoplasmic and Nuclear
;
Receptors, Steroid
;
Repressor Proteins
;
Saccharomyces cerevisiae/*genetics
;
*Saccharomyces cerevisiae Proteins
;
*Serine Endopeptidases
;
Transcription Factors/genetics/*metabolism/pharmacology
;
Transcription, Genetic
;
Transfection
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
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