title: Deletion in the promoter region and altered expression of Pitx3 homeobox gene in aphakia mice.
abstract: Mouse aphakia (ak) is a recessive phenotype that spontaneously occurs in the 129/Sv-SlJ strain and is characterized by small eyes that lack a lens. We have recently identified a homeobox-containing gene, Pitx3, and have shown that it is expressed in the developing lens and maps to chromosome 19 close to ak in mouse. Human PITX3 gene was found to underlie anterior segment dysgenesis and cataracts. We have now obtained the entire sequence of the mouse Pitx3 gene including 10 kb of the 5' region and 5 kb of the 3' region. Of several microsatellite repeat regions identified within the Pitx3 sequence, one was informative for linkage analysis. No recombination was observed between ak and the Pitx3 marker, indicating that these two loci are closely linked (0.2 +/- 0.2 cM). Additionally, Pitx3 transcripts were not detected in the ak/ak mice either in the lens placode or at later developmental stages of the lens by in situ hybridization. Since no differences were previously found between ak/ak and wild-type sequences in the Pitx3 coding region, we hypothesized that an etiologic mutation is located in the promoter or other regulatory regions. To test this hypothesis we studied the 5' flanking region of the Pitx3 gene. This analysis revealed a deletion of 652 bp located 2.5 kb upstream from the start point of the Pitx3 5' UTR sequence in ak/ak mice. The deletion co-segregated with the ak mutation and was not detected in 16 samples from 10 different mouse strains including the founder strains. Analysis of the 652 bp region identified sequences similar to consensus binding sites for transcription factors AP-2 and Maf that were shown to play a critical role in lens determination. These lines of evidence suggest that the abnormal ocular development in the aphakia mouse is due to the deletion upstream of the Pitx3 gene.
authors.affiliation: Department of Pediatrics, University of Iowa, Iowa City 52242, USA. esemina@blue.weeg.uiowa.edu
authors.forename: E V
authors.initials: EV
authors.lastname: Semina
authors.forename: J C
authors.initials: JC
authors.lastname: Murray
authors.forename: R
authors.initials: R
authors.lastname: Reiter
authors.forename: R F
authors.initials: RF
authors.lastname: Hrstka
authors.forename: J
authors.initials: J
authors.lastname: Graw
mesh_terms: D000123:Acetyltransferases; D000818:Animals; D001035:Aphakia / Q000235:genetics* / Q000473:pathology; D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000235:genetics; D004622:Embryo, Mammalian / Q000002:abnormalities / Q000378:metabolism; D000081031:Fatty Acid Elongases; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005801:Genes, Homeobox / Q000235:genetics; D008040:Genetic Linkage; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics*; D017403:In Situ Hybridization; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D008806:Mice, Inbred AKR; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008815:Mice, Inbred Strains; D008817:Mice, Mutant Strains; D008969:Molecular Sequence Data; D009115:Muridae; D009687:Nuclear Proteins; D051761:Paired Box Transcription Factors; D011401:Promoter Regions, Genetic / Q000235:genetics*; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D014157:Transcription Factors / Q000235:genetics*; D000097577:Homeobox Protein PITX2
chemical_list: D000123:Acetyltransferases; D004247:DNA; D000081031:Fatty Acid Elongases; D020662:Guanine Nucleotide Exchange Factors; D018398:Homeodomain Proteins; D008565:Membrane Proteins; D009687:Nuclear Proteins; D051761:Paired Box Transcription Factors; D014157:Transcription Factors; D000097577:Homeobox Protein PITX2; C120445:GBF1 protein, human; C101485:homeobox protein PITX1; C433629:homeobox protein PITX3