Enhancer-independent promoter activity of the mouse αB-crystallin/small heat shock protein gene in the lens and cornea of transgenic mice

• Published in: Mechanisms of development 2000-04, Vol.92 (2), p.125-134
• Main Authors: Gopal-Srivastava, Rashmi, Kays, W.Todd, Piatigorsky, Joram
• Format: Article
• Language: English
• Subjects: Cornea; Crystallin; Enhancer; Gene expression; Lens; Promoter activity; Small heat shock protein
• ISSN: 0925-4773; 1872-6356
• DOI: 10.1016/S0925-4773(99)00341-X

The αB-crystallin/small heat shock protein gene is expressed very highly in the mouse eye lens and to a lesser extent in many other nonocular tissues, including the heart, skeletal muscle and brain. Previously we showed in transgenic mice that lens-specific αB-crystallin promoter activity is directed by a proximal promoter fragment (−164/+44) and that non-lens promoter activity depends on an upstream enhancer (−427/−259) composed of at least 5 cis-control elements. Here we have used truncated αB-crystallin promoter-CAT transgenes to test by biphasic CAT assays and/or histochemistry for specific expression in the cornea and lens. Deletion either of 87 bp (−427/−340) from the 5′ end of the αB-crystallin enhancer or of the whole enhancer (−427/−258) abolished αB-crystallin promoter activity in all tissues except the lens and corneal epithelium when examined by the biphasic CAT assay in 4–5-week-old transgenic mice. These truncations also lowered promoter strength in the lens. The −426/+44-CAT, −339/+44-CAT and −164/+44-CAT (previously thought to be lens-specific in transgenic mice) transgenes were all expressed in the 4–6-week-old corneal epithelium when examined histochemically. Immunohistochemical staining confirmed the presence of endogenous αB-crystallin in the mature corneal epithelial cells. CAT gene expression driven by the αB-crystallin promoter with or without the enhancer was evident in the embryonic and 4–6-week-old lens. By contrast, activity of the αB-crystallin promoter/enhancer-CAT transgene was not detectable in the corneal epithelium before birth. Taken together, these results indicate that the intact enhancer of the αB-crystallin/small heat shock protein gene is required for promoter activity in all tissues tested except the lens and cornea.