Overexpression of PAX6(5a) in lens fiber cells results in cataract and upregulation of (alpha)5(beta)1 integrin expression.
The PAX6 gene, a key regulator of eye development, produces two major proteins that differ in paired domain structure: PAX6 and PAX6(5a). It is known that an increase in the PAX6(5a) to PAX6 ratio leads to multiple ocular defects in humans. Here, transgenic mice were created that overexpress human PAX6(5a) in the lens. These mice develop cataracts with abnormalities in fiber cell shape as well as fiber cell/lens capsule and fiber cell/fiber cell interactions. While the structure of the actin cytoskeleton appeared relatively normal, the cataractous lens expresses increased amounts of paxillin and p120(ctn) as well as large aggregates of (alpha)5(beta)1 integrin in the dysgenic fiber cells. The elevated amounts of these proteins in the transgenic lens correlated well with elevated levels of their respective mRNAs. To investigate the role of Pax-6(5a) in the upregulation of these genes, a series of gel shift experiments using truncated proteins and consensus oligonucleotides demonstrated the complexity of Pax-6 and Pax-6(5a) binding to DNA, aiding our identification of potential binding sites in the human (&agr;)5- and (beta)1-integrin promoters. Consequent gel shift analysis demonstrated that these putative regulatory sequences bind Pax-6 and/or Pax-6(5a) in lens nuclear extracts, suggesting that the human (alpha)5 and (beta)1 integrin promoters contain PAX6/PAX6(5a) binding sites and maybe directly regulated by this transcription factor in the transgenic lens. We conclude that these transgenic mice are good models to study a type of human cataract and for identifying batteries of genes that are directly or indirectly regulated by both forms of Pax-6.
Duncan
M K
MK
Department of Biological Sciences, University of Delaware, Newark, DE 19716, USA. duncanm@udel.edu
Kozmik
Z
Z
Cveklova
K
K
Piatigorsky
J
J
Cvekl
A
A
D000818:Animals; D015703:Antigens, CD / Q000235:genetics*; D001665:Binding Sites; D002386:Cataract / Q000209:etiology / Q000378:metabolism*; D015815:Cell Adhesion Molecules / Q000096:biosynthesis; D003459:Crystallins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D003598:Cytoskeletal Proteins / Q000235:genetics; D004247:DNA / Q000378:metabolism; D005136:Eye Proteins; D015870:Gene Expression; D018398:Homeodomain Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D006801:Humans; D039482:Integrin alpha5; D019012:Integrin beta1 / Q000235:genetics*; D007908:Lens, Crystalline / Q000473:pathology*; D051379:Mice; D008822:Mice, Transgenic; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; D051419:Paxillin; D010750:Phosphoproteins / Q000235:genetics; D020033:Protein Isoforms / Q000235:genetics / Q000502:physiology; D012333:RNA, Messenger; D012097:Repressor Proteins; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D015854:Up-Regulation
D015703:Antigens, CD; D015815:Cell Adhesion Molecules; D003459:Crystallins; D003598:Cytoskeletal Proteins; D005136:Eye Proteins; D018398:Homeodomain Proteins; D039482:Integrin alpha5; D019012:Integrin beta1; D000071841:PAX6 Transcription Factor; C000605730:PAX6 protein, human; C496061:PXN protein, human; D051761:Paired Box Transcription Factors; C000605732:Pax6 protein, mouse; D051419:Paxillin; D010750:Phosphoproteins; D020033:Protein Isoforms; C496062:Pxn protein, mouse; D012333:RNA, Messenger; D012097:Repressor Proteins; D014157:Transcription Factors; D004247:DNA