TY - JOUR
T1 - Pax-6 and αB-crystallin/small heat shock protein gene regulation in the murine lens. Interaction with the lens-specific regions, LSR1 and LSR2
AU - Gopal-Srivastava, Rashmi
AU - Cvekl, Ales
AU - Piatigorsky, Joram
PY - 1996
Y1 - 1996
N2 - We have demonstrated previously that a transgene comprising the - 164/+44 fragment of the murine αB-crystallin gene fused to the bacterial chloramphenicol acetyltransferase (cat) gene is lens-specific in transgenic mice. The -147 to -118 sequence was identified as a lens-specific regulatory region and is called here LSR1 for lens-specific region 1. In the present experiments, a -115/+44-cat transgene was also lens-specific in transgenic mice, although the average activity was 30 times lower than that derived from the -164/+44-cat transgene. The -115/+44 αB-crystallin fragment contains a highly conserved region (-78 to -46) termed here LSR2. A -68/+44-cat transgene, in which LSR2 is truncated, was inactive in transgenic mice. DNase I footprinting indicated that LSR1 and LSR2 bind partially purified nuclear proteins from either αTN4-1 lens cells or the mouse lens as well as the purified paired domain of Pax-6. Site-specific mutation of LSR1 eliminated both Pax-6 binding and promoter activity of the -164/+ 44-cat transgene in transgenic mice. Finally antibody/electrophoretic mobility shift assays and cotransfection experiments indicated that Pax-6 can activate the αB- crystallin promoter via LSR1 and LSR2. Our data strengthen the idea that Pax- 6 has had a major role in recruiting genes for high expression in the lens.
AB - We have demonstrated previously that a transgene comprising the - 164/+44 fragment of the murine αB-crystallin gene fused to the bacterial chloramphenicol acetyltransferase (cat) gene is lens-specific in transgenic mice. The -147 to -118 sequence was identified as a lens-specific regulatory region and is called here LSR1 for lens-specific region 1. In the present experiments, a -115/+44-cat transgene was also lens-specific in transgenic mice, although the average activity was 30 times lower than that derived from the -164/+44-cat transgene. The -115/+44 αB-crystallin fragment contains a highly conserved region (-78 to -46) termed here LSR2. A -68/+44-cat transgene, in which LSR2 is truncated, was inactive in transgenic mice. DNase I footprinting indicated that LSR1 and LSR2 bind partially purified nuclear proteins from either αTN4-1 lens cells or the mouse lens as well as the purified paired domain of Pax-6. Site-specific mutation of LSR1 eliminated both Pax-6 binding and promoter activity of the -164/+ 44-cat transgene in transgenic mice. Finally antibody/electrophoretic mobility shift assays and cotransfection experiments indicated that Pax-6 can activate the αB- crystallin promoter via LSR1 and LSR2. Our data strengthen the idea that Pax- 6 has had a major role in recruiting genes for high expression in the lens.
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U2 - 10.1074/jbc.271.38.23029
DO - 10.1074/jbc.271.38.23029
M3 - Article
C2 - 8798491
AN - SCOPUS:0029661444
SN - 0021-9258
VL - 271
SP - 23029
EP - 23036
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 38
ER -