title: Tissue-specific expression of the chicken alpha A-crystallin gene in cultured lens epithelia and transgenic mice.
abstract: The present experiments show that the single gene for the lens-specific protein alpha A-crystallin of chickens and mice uses a different subset of cis- and trans-acting regulatory elements for expression in transfected embryonic chicken lens epithelial cells. A chicken alpha A-crystallin-chloramphenicol acetyltransferase (CAT) fusion gene required 162 base pairs whereas the murine alpha A-crystallin-CAT fusion gene required only 111 base pairs of 5'-flanking sequences for efficient tissue-specific expression in the transfected chicken lens cells. Gel retardation and competition experiments were performed using embryonic chicken lens nuclear extract and oligodeoxynucleotides identical to the 5'-flanking region of the chicken (-170/-111) and murine (-111/-88 and -88/-55) alpha A-crystallin gene. The results indicated that these homologous promoters use different nuclear factors for function. Methylation interference analysis identified a dyad of symmetry (CTGGTTCCCACCAG) at position -153 to -140 in the chicken alpha A-crystallin promoter which binds one or more lens nuclear factors. Gel mobility shift experiments using nuclear extracts of brain, reticulocytes, and muscle of embryonic chickens or HeLa cells suggested that the factor(s) binding to the chicken alpha A-crystallin gene promoter sequences are not lens specific. Despite differences in the functional and protein-binding properties of the alpha A-crystallin gene promoter of chickens and mice, expression of the chicken alpha A-crystallin-CAT fusion gene in transgenic mice was lens specific, consistent with a common underlying mechanism for expression of the alpha A-crystallin gene in chickens and mice.
authors.affiliation: Laboratory of Molecular and Developmental Biology, National Eye Institute, Bethesda, Maryland 20892.
authors.forename: J F
authors.initials: JF
authors.lastname: Klement
authors.forename: E F
authors.initials: EF
authors.lastname: Wawrousek
authors.forename: J
authors.initials: J
authors.lastname: Piatigorsky
mesh_terms: D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002645:Chickens; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003459:Crystallins / Q000235:genetics*; D004848:Epithelium / Q000378:metabolism; D015870:Gene Expression; D005796:Genes; D007908:Lens, Crystalline / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000138:chemical synthesis; D010957:Plasmids; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
chemical_list: D003459:Crystallins; D015345:Oligonucleotide Probes; D015500:Chloramphenicol O-Acetyltransferase