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J. Biol. Chem., Vol. 279, Issue 30, 31348-31356, July 23, 2004

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Neurokinin-B Transcription in Erythroid Cells

DIRECT ACTIVATION BY THE HEMATOPOIETIC TRANSCRIPTION FACTOR GATA-1^*

Saumen Pal, Michael J. Nemeth, David Bodine, Jeffery L. Miller¶, John Svaren||, Swee Lay Thein**, Philip J. Lowry, and Emery H. Bresnick

From the University of Wisconsin Medical School, Molecular and Cellular Pharmacology Program, Department of Pharmacology, Madison, Wisconsin 53706, NHGI, National Institutes of Health, Bethesda, Maryland 20892, ^¶NIDDK, National Institutes of Health, Bethesda, Maryland 20892, ^**Hematological Medicine, King's Denmark Hill Campus, London SE5 9PJ, United Kingdom, ^||University of Wisconsin, Department of Comparative Biosciences, Madison, Wisconsin 53706, and School of Animal and Microbial Sciences, University of Reading, Reading RG6 6AJ, United Kingdom

The GATA family of transcription factors establishes genetic networks that control developmental processes including hematopoiesis, vasculogenesis, and cardiogenesis. We found that GATA-1 strongly activates transcription of the Tac-2 gene, which encodes proneurokinin-B, a precursor of neurokinin-B (NK-B). Neurokinins function through G protein-coupled transmembrane receptors to mediate diverse physiological responses including pain perception and the control of vascular tone. Whereas an elevated level of NK-B was implicated in pregnancy-associated pre-eclampsia (Page, N. M., Woods, R. J., Gardiner, S. M., Lomthaisong, K., Gladwell, R. T., Butlin, D. J., Manyonda, I. T., and Lowry, P. J. (2000) Nature 405, 797–800), the regulation of NK-B synthesis and function are poorly understood. Tac-2 was expressed in normal murine erythroid cells and was induced upon ex vivo erythropoiesis. An estrogen receptor fusion to GATA-1 (ER-GATA-1) and endogenous GATA-1 both occupied a region of Tac-2 intron-7, which contains two conserved GATA motifs. Genetic complementation analysis in GATA-1-null G1E cells revealed that endogenous GATA-2 occupied the same region of intron-7, and expression of ER-GATA-1 displaced GATA-2 and activated Tac-2 transcription. Erythroid cells did not express neurokinin receptors, whereas aortic and yolk sac endothelial cells differentially expressed neurokinin receptor subtypes. Since NK-B induced cAMP accumulation in yolk sac endothelial cells, these results suggest a new mode of vascular regulation in which GATA-1 controls NK-B synthesis in erythroid cells.

Received for publication, March 29, 2004

^* This work was supported in part by National Institutes of Health Grants DK50107 and DK55700 (to E. H. B.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

A Romnes Scholar and a Shaw Scientist. To whom correspondence should be addressed: University of Wisconsin Medical School, Molecular and Cellular Pharmacology Program, Dept. of Pharmacology, 1300 University Ave., 383 Medical Sciences Center, Madison, WI 53706. Tel.: 608-265-6446; Fax: 608-262-1257; E-mail: ehbresni{at}wisc.edu.

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