Pax-6 and αB-crystallin/small heat shock protein gene regulation in the murine lens. Interaction with the lens-specific regions, LSR1 and LSR2

Rashmi Gopal-Srivastava, Ales Cvekl, Joram Piatigorsky

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish (US)
Pages (from-to)23029-23036
Number of pages8
JournalJournal of Biological Chemistry
Volume271
Issue number38
DOIs
StatePublished - 1996
Externally publishedYes

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Small Heat-Shock Proteins
Crystallins
Gene expression
Lenses
Chloramphenicol O-Acetyltransferase
Transgenes
Transgenic Mice
Genes
Electrophoretic mobility
Nucleic Acid Regulatory Sequences
Deoxyribonuclease I
Electrophoretic Mobility Shift Assay
Nuclear Proteins
Assays
Experiments
Gene Expression
Mutation
Antibodies

ASJC Scopus subject areas

  • Biochemistry

Cite this

Pax-6 and αB-crystallin/small heat shock protein gene regulation in the murine lens. Interaction with the lens-specific regions, LSR1 and LSR2. / Gopal-Srivastava, Rashmi; Cvekl, Ales; Piatigorsky, Joram.

In: Journal of Biological Chemistry, Vol. 271, No. 38, 1996, p. 23029-23036.

Research output: Contribution to journalArticle

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