The Drosophila nuclear receptors FTZ-F1 alpha and FTZ-F1 beta compete as monomers for binding to a site in the fushi tarazu gene.

The striped pattern of fushi tarazu (ftz) expression found in the blastoderm of the Drosophila melanogaster embryo is generated largely through complex interactions between multiple transcription factors that bind to the zebra element of the ftz gene. A motif in the zebra element, the FTZ-F1 recognition element (F1RE), has been shown to bind a transcription factor, FTZ-F1 alpha, that is a member of the nuclear receptor family. We recently identified a second, related member of this family, FTZ-F1 beta, that also binds to this motif. To investigate the possibility that FTZ-F1 alpha and FTZ-F1 beta coregulate ftz transcription through the F1RE, we have studied the DNA binding properties of FTZ-F1 alpha and FTZ-F1 beta. We demonstrate that recombinant FTZ-F1 alpha and FTZ-F1 beta proteins produce similar in vitro DNase I footprint patterns on a 14-nucleotide region of the zebra element and bind to this site with similar affinities and sequence specificities. Using wild-type and N-terminally truncated receptors, we have determined that FTZ-F1 alpha and FTZ-F1 beta both bind as monomers to the 9-bp F1RE in the zebra element, as well as to an imperfect inverted F1RE repeat present in the Drosophila alcohol dehydrogenase gene. A polyclonal antibody raised against FTZ-F1 beta identifies a predominant F1RE-binding component in embryonic nuclear extracts. Although FTZ-F1 alpha is also present in these extracts, FTZ-F1 alpha and FTZ-F1 beta do not appear to form heterodimers with each other. Cotransfection assays in mammalian cell culture indicate that both receptors contribute to the net transcriptional activity of a reporter gene through their direct interaction with the F1RE. These data suggest that FTZ-F1 alpha and FTZ-F1 beta likely coregulate common target genes by competition for binding to a 9-bp recognition element.

New retinoid X receptor subtypes in zebra fish (Danio rerio) differentially modulate transcription and do not bind 9-cis retinoic acid.

Retinoid X receptors (RXRs), along with retinoic acid (RA) receptors (RARs), mediate the effects of RA on gene expression. Three subtypes of RXRs (alpha, beta, and gamma) which bind to and are activated by the 9-cis stereoisomer of RA have been characterized. They activate gene transcription by binding to specific sites on DNA as homodimers or as heterodimers with RARs and other related nuclear receptors, including the vitamin D receptor, thyroid hormone receptors (TRs), and peroxisome proliferator-activated receptors. Two additional RXR subtypes (delta and epsilon) isolated from zebra fish cDNA libraries are described here; although both subtypes form DNA-binding heterodimers with RARs and TR, neither binds 9-cis RA, and both are transcriptionally inactive on RXR response elements. In cotransfection studies with TR, the delta subtype was found to function in a dominant negative manner, while the epsilon subtype had a slight stimulatory effect on thyroid hormone (T3)-dependent transcriptional activity. The discovery of these two novel receptors in zebra fish expands the functional repertoire of RXRs to include ligand-independent and dominant negative modulation of type II receptor function.

FTZ-F1 beta, a novel member of the Drosophila nuclear receptor family.

The Drosophila melanogaster gene FTZ-F1 beta, encoding a novel member of the steroid/thyroid hormone receptor gene superfamily, was isolated by cross-hybridization with a complementary DNA for the Drosophila nuclear receptor, FTZ-F1 (Lavorgna et al., 1991). The cDNA deduced protein sequence for FTZ-F1 beta displays significant amino acid identity with other vertebrate and invertebrate nuclear receptors, most notably with FTZ-F1. Also, bacterially expressed FTZ-F1 beta protein binds to a FTZ-F1 binding site found in the zebra stripe promoter element of the segmentation gene fushi tarazu (ftz). Northern blot analysis detected FTZ-F1 beta expression at all stages of the Drosophila life cycle including a possible maternal component. In situ hybridization in whole-mounted embryos localized transcripts for FTZ-F1 beta evenly expressed throughout the blastodermal layer in early embryos. At later stages of development strong FTZ-F1 beta expression is observed in both the brain and ventral chord structures as well as in the hindgut. Temporal and spatial expression patterns of the FTZ-F1 beta gene suggest that it may have multiple roles in early embryogenesis, neurogenesis, and in the adult. Furthermore, the identification of FTZ-F1 beta as a nuclear receptor family member suggests that an as yet undiscovered FTZ-F1 beta specific ligand is involved in Drosophila development.