Developmental regulation of the chicken beta B1-crystallin promoter in transgenic mice.
The cis-elements responsible for the high-level, lens-specific expression of the chicken beta B1-crystallin gene were investigated by generating mice harboring beta B1-crystallin promoter/chloramphenicol acetyl transferase (CAT) transgenes. Deletion of promoter sequences -434/-153 and -152/-127 as well as site-directed mutagenesis of the PL1 (-116/-102) and Pl2 (-90/-76) elements significantly decreased CAT gene expression in the lenses of adult transgenic mice. Transfection studies using multimerized PL1 and PL2 elements fused to the chicken beta-actin basal promoter indicated that PL1 is a general activating element while PL2 is involved in the lens-specificity of the chicken beta B1-crystallin promoter. CAT histochemistry demonstrated that the chicken beta B1-crystallin promoter (-434/+30) was active in both primary and secondary lens fiber cells from 12.5 days post coitum (dpc) until adulthood. Activity of the -152/+30/CAT transgene was relatively low and confined to the primary lens fiber cells of 16.5 dpc mice. Together, these data suggest that the reduced activity of this promoter in the adult lens is due both to this developmentally restricted expression pattern and a reduction in promoter activity. RNA hybridization studies demonstrated that the chicken beta B1-crystallin/CAT (-434/+30) transgene was expressed at similar levels in the same cells as the endogenous mouse beta B1-crystallin gene in 16.5 dpc transgenic mouse embryos. These data show a strict conservation of the lens-specific spatial and temporal regulation of the chicken and mouse beta B1-crystallin genes.
Duncan
M K
MK
Laboratory of Molecular and Developmental Biology, National Eye Institute/National Institutes of Health, Bethesda, MD 20892, USA.
Li
X
X
Ogino
H
H
Yasuda
K
K
Piatigorsky
J
J
D000367:Age Factors; D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D015139:Blotting, Southern; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003459:Crystallins / Q000235:genetics*; D004622:Embryo, Mammalian; D018507:Gene Expression Regulation, Developmental; D017403:In Situ Hybridization; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic; D014018:Tissue Distribution; D014162:Transfection
D003459:Crystallins; D015500:Chloramphenicol O-Acetyltransferase