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Spatially specific expression of Hoxb4 is dependent on the ubiquitous transcription factor NFY

Jonathan Gilthorpe^1,*,¶, Marie Vandromme^1,,¶, Tim Brend^1,, Alejandro Gutman¹, Dennis Summerbell^1,, Nick Totty^2, and Peter W. J. Rigby^1,,**

¹ Division of Eukaryotic Molecular Genetics, MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK
² Ludwig Institute for Cancer Research, 91 Riding House Street, London W1W 7BS, UK
^* Present address: Department of Developmental Neurobiology, King’s College London, Guy’s Campus, London Bridge, London SE1 1UL, UK
Present address: Laboratoire de Biologie Moleculaire et Cellulaire, UMR 5665 CNRS/ENS Lyon, 46 Allee d’Italie, 69364 Lyon Cedex 07, France
Present address: Section of Gene Function and Regulation, The Institute of Cancer Research, Chester Beatty Laboratories, 237 Fulham Road, London SW3 6JB, UK
Present address: Protein Analysis Laboratory, Cancer Research UK, PO Box123, Lincoln’s Inn Fields, London WC2A 3PX, UK
^¶ These authors contributed equally to this article and should both be considered first authors

**Author for correspondence (e-mail: p.rigby{at}icr.ac.uk)

Accepted 22 May 2002

Understanding how boundaries and domains of Hox gene expression are determined is critical to elucidating the means by which the embryo is patterned along the anteroposterior axis. We have performed a detailed analysis of the mouse Hoxb4 intron enhancer to identify upstream transcriptional regulators. In the context of an heterologous promoter, this enhancer can establish the appropriate anterior boundary of mesodermal expression but is unable to maintain it, showing that a specific interaction with its own promoter is important for maintenance. Enhancer function depends on a motif that contains overlapping binding sites for the transcription factors NFY and YY1. Specific mutations that either abolish or reduce NFY binding show that it is crucial for enhancer activity. The NFY/YY1 motif is reiterated in the Hoxb4 promoter and is known to be required for its activity. As these two factors are able to mediate opposing transcriptional effects by reorganizing the local chromatin environment, the relative levels of NFY and YY1 binding could represent a mechanism for balancing activation and repression of Hoxb4 through the same site.

Key words: Hoxb4, Transcriptional regulation, NFY, YY1, Embryo, Transcription factor, Mouse

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