Comparative genomics reveals functional transcriptional control sequences in the Prop1 gene.
Mutations in PROP1 are a common genetic cause of multiple pituitary hormone deficiency (MPHD). We used a comparative genomics approach to predict the transcriptional regulatory domains of Prop1 and tested them in cell culture and mice. A BAC transgene containing Prop1 completely rescues the Prop1 mutant phenotype, demonstrating that the regulatory elements necessary for proper PROP1 transcription are contained within the BAC. We generated DNA sequences from the PROP1 genes in lemur, pig, and five different primate species. Comparison of these with available human and mouse PROP1 sequences identified three putative regulatory sequences that are highly conserved. These are located in the PROP1 promoter proximal region, within the first intron of PROP1, and downstream of PROP1. Each of the conserved elements elicited orientation-specific enhancer activity in the context of the Drosophila alcohol dehydrogenase minimal promoter in both heterologous and pituitary-derived cells lines. The intronic element is sufficient to confer dorsal expansion of the pituitary expression domain of a transgene, suggesting that this element is important for the normal spatial expression of endogenous Prop1 during pituitary development. This study illustrates the usefulness of a comparative genomics approach in the identification of regulatory elements that may be the site of mutations responsible for some cases of MPHD.
Ward
Robert D
RD
Graduate Program in Cellular and Molecular Biology, University of Michigan, Ann Arbor, Michigan, USA.
Davis
Shannon W
SW
Cho
Minchul
M
Esposito
Constance
C
Lyons
Robert H
RH
Cheng
Jan-Fang
JF
Rubin
Edward M
EM
Rhodes
Simon J
SJ
Raetzman
Lori T
LT
Smith
Timothy P L
TP
Camper
Sally A
SA
D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D017124:Conserved Sequence; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D018507:Gene Expression Regulation, Developmental; D023281:Genomics; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D010902:Pituitary Gland / Q000196:embryology; D012045:Regulatory Sequences, Nucleic Acid / Q000502:physiology*; D017386:Sequence Homology, Amino Acid
D018398:Homeodomain Proteins; C102134:Prophet of Pit-1 protein