Identification of target genes regulated by homeotic proteins in Drosophila melanogaster through genetic selection of Ultrabithorax protein-binding sites in yeast.
A method based on the transcriptional activation of a selectable reporter in yeast cells was used to identify genes regulated by the Ultrabithorax homeoproteins in Drosophila melanogaster. Fifty-three DNA fragments that can mediate activation by UBX isoform Ia in this test were recovered after screening 15% of the Drosophila genome. Half of these fragments represent single-copy sequences in the genome. Six single-copy fragments were investigated in detail, and each was found to reside near a transcription unit whose expression in the embryo is segmentally modulated as expected for targets of homoeotic genes. Four of these putative target genes are expressed in patterns that suggest roles in the development of regional specializations within mesoderm derivatives; in three cases these expression patterns depend on Ultrabithorax function. Extrapolation from this pilot study indicates that 85-170 candidate target genes can be identified by screening the entire Drosophila genome with UBX isoform Ia. With appropriate modifications, this approach should be applicable to other transcriptional regulators in diverse organisms.
Mastick
G S
GS
Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213.
McKay
R
R
Oligino
T
T
Donovan
K
K
López
A J
AJ
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004064:Digestive System / Q000196:embryology / Q000378:metabolism; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000196:embryology / Q000235:genetics*; D018628:Gene Dosage; D005786:Gene Expression Regulation; D017344:Genes, Insect / Q000235:genetics*; D017930:Genes, Reporter; D006066:Gonads / Q000196:embryology / Q000378:metabolism; D006257:Head / Q000196:embryology / Q000502:physiology; D018398:Homeodomain Proteins / Q000378:metabolism*; D008648:Mesoderm / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012641:Selection, Genetic; D012689:Sequence Homology, Nucleic Acid; D013623:Tail / Q000196:embryology / Q000378:metabolism; D014157:Transcription Factors; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D018398:Homeodomain Proteins; D014157:Transcription Factors; C066177:Ubx protein, Drosophila