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Volume 272, Number 4, Issue of January 24, 1997 pp. 2167-2175
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

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Retinoic Acid-responsive Enhancers Located 3 of the Hox A and Hox B Homeobox Gene Clusters
FUNCTIONAL ANALYSIS

(Received for publication, July 16, 1996, and in revised form, October 18, 1996)

From the Department of Pharmacology, Cornell University Medical College, New York, New York 10021

Homeobox genes control the spatial identity and differentiation of tissues in the developing vertebrate embryo. Retinoids are signaling molecules involved in the regulation of Hox genes. We previously identified a 3 enhancer called the RAIDR₅, which contained a DR5 retinoic acid response element (RARE) and was responsible for the retinoic acid (RA)-associated expression of the murine Hoxa-1 gene in teratocarcinoma cells. We demonstrate that a similar enhancer, which contains a DR₅ RARE, is located at a DNase I-hypersensitive site 3 of the murine Hoxb-1 gene. This enhancer, the Hoxb-1 RAIDR₅, regulates the RA responsiveness of the Hoxb-1 gene and is different in location and sequence from the RA-regulated 3 Hoxb-1 enhancers previously described. Several DNA elements within the murine Hoxa-1 RA-inducible RAIDR₅ enhancer, including the DR₅ RARE, conserved element (CE) 1, and CE2, are conserved in the murine Hoxb-1 RAIDR₅ enhancer, the human homolog of Hoxa-1, and in the chicken Hoxb-1 gene. Gel shifts show that the CE2 sequence TATTTACTCA binds an RA-inducible factor, while UV cross-linking indicates that a 170-kDa protein binds to this sequence. Thus, the Hoxa-1 and Hoxb-1 genes possess 3 enhancers with similar sequences through which their expression and responsiveness to endogenous retinoids are controlled.

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