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Science 31 May 1991:
Vol. 252. no. 5010, pp. 1296 - 1300
DOI: 10.1126/science.1925541
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Articles

Science, Vol 252, Issue 5010, 1296-1300
Copyright © 1991 by American Association for the Advancement of Science

articles

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

TE Wilson, TJ Fahrner, M Johnston, and J Milbrandt

Department of Pathology, Washington University School of Medicine, St. Louis, MO 63110.

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.


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Endocrinology 138, 4138-4146
   Abstract »    Full Text »    PDF »
The orphan nuclear receptor LXRalpha is positively and negatively regulated by distinct products of mevalonate metabolism.
B. M. Forman, B. Ruan, J. Chen, G. J. Schroepfer Jr., and R. M. Evans (1997)
PNAS 94, 10588-10593
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Multiple Orphan Nuclear Receptors Converge to Regulate Rat P450c17 Gene Transcription: Novel Mechanisms for Orphan Nuclear Receptor Action.
P. Zhang and S. H. Mellon (1997)
Mol. Endocrinol. 11, 891-904
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The DNA Binding Pattern of the Retinoid X Receptor Is Regulated by Ligand-dependent Modulation of Its Oligomeric State.
S. Kersten, H. Gronemeyer, and N. Noy (1997)
J. Biol. Chem. 272, 12771-12777
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Cloning and Characterization of a Transcription Factor That Binds to the Proximal Promoters of the Two Mouse Type I Collagen Genes.
T. Hasegawa, A. Takeuchi, O. Miyaishi, K.-i. Isobe, and B. de Crombrugghe (1997)
J. Biol. Chem. 272, 4915-4923
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Recombinant Microorganisms as Tools for High Throughput Screening for Nonantibiotic Compounds.
R. D. Klein and T. G. Geary (1997)
J Biomol Screen 2, 41-49
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Induction of the Intronic Enhancer of the Human Ciliary Neurotrophic Factor Receptor (CNTFRalpha ) Gene by the TR4 Orphan Receptor. A MEMBER OF STEROID RECEPTOR SUPERFAMILY.
W.-J. Young, S. M. Smith, and C. Chang (1997)
J. Biol. Chem. 272, 3109-3116
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