Plasticity of Drosophila paired function.
The Drosophila Paired (Prd) transcription factor has homeodomain (HD) and paired domain (PD) DNA-binding activities required for in vivo function. Correspondingly, Prd activation of late even-skipped (eve) expression occurs through a conserved target sequence (PTE) with HD and PD half sites, both of which are required for activation. To investigate the relationship between the HD and PD, and their roles in conferring specificity to Prd function, we tested altered versions of the Prd protein and of the PTE target site using in vivo assays in embryos. We found that function through PTE was constrained by the targeting specifications of both the HD and PD as well as the spatial relationship between these two domains. PTE function was also constrained by the spacing between the target half sites for the PD and HD, although surprisingly, late eve activation was retained when PTE was replaced by in vitro optimized binding sites for either the PD alone or for an HD dimer. In contrast to late eve regulation, other Prd targets tolerated more changes in the Prd protein, suggesting that their target sequences may be qualitatively different from PTE.
Department of Biology, Wesleyan University, Middletown, Connecticut 06459, USA.
Y
Y
Lan
M
M
Fujioka
R
R
Polsgrove
P
P
Miskiewicz
D
D
Morrissey
T
T
Goto
M
M
Weir
D000818:Animals; D030801:Animals, Genetically Modified; D001426:Bacterial Proteins; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D019521:Body Patterning / Q000235:genetics*; D004247:DNA / Q000235:genetics / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D004330:Drosophila / Q000196:embryology* / Q000235:genetics*; D029721:Drosophila Proteins; D018507:Gene Expression Regulation, Developmental; D005801:Genes, Homeobox; D017344:Genes, Insect; D018398:Homeodomain Proteins / Q000235:genetics*; D016296:Mutagenesis; D014157:Transcription Factors
D001426:Bacterial Proteins; D017931:DNA Primers; D029721:Drosophila Proteins; D018398:Homeodomain Proteins; D014157:Transcription Factors; C052837:eve protein, Drosophila; C489935:prd protein, Drosophila; D004247:DNA