Identification of the DNA binding site for NGFI-B by genetic selection in yeast

Thomas E. Wilson, Timothy J. Fahrner, Mark Johnston, Jeffrey Milbrandt

Research output: Contribution to journalArticle

456 Scopus citations

Abstract

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 functionaily 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.

Original languageEnglish
Pages (from-to)1296-1300
Number of pages5
JournalScience
Volume252
Issue number5010
DOIs
StatePublished - Jan 1 1991

Fingerprint Dive into the research topics of 'Identification of the DNA binding site for NGFI-B by genetic selection in yeast'. Together they form a unique fingerprint.

  • Cite this