Distinct embryonic expression and localization of CBP and p300 histone acetyltransferases at the mouse alphaA-crystallin locus in lens.

Mouse alphaA-crystallin gene encodes the most abundant protein of the mammalian lens. Expression of alphaA-crystallin is regulated temporally and spatially during lens development with initial expression in the lens vesicle followed by strong upregulation in the differentiating primary lens fibers. Lens-specific expression of alphaA-crystallin is mediated by DNA-binding transcription factors Pax6, c-Maf and CREB bound to its promoter region. Its 5'-distal enhancer, DCR1, mediates regulation of alphaA-crystallin via FGF signaling, while its 3'-distal enhancer, DCR3, functions only in elongated primary lens fibers via other lens differentiation pathways. DCR1 and DCR3 establish outside borders of a lens-specific chromatin region marked by histone H3 K9 acetylation. Here, we identified CREB-binding protein (CBP) and p300 as major histone acetyltransferases (HATs) associated in vivo with the mouse alphaA-crystallin locus. Both HATs are expressed in embryonic lens. Expression of CBP in primary lens fiber cells coincides with alphaA-crystallin. In the chromatin of lens epithelial cells, chromatin immunoprecipitations (ChIPs) show that the alphaA-crystallin promoter is notably devoid of any significant presence of CBP and p300, though DCR1 and a few other regions show the presence of these HATs. In the chromatin obtained from newborn lens, CBP was localized specifically at the promoter region with about ten times higher abundance compared to the entire alphaA-crystallin locus. In contrast, p300 is distributed more evenly across the entire locus. Analysis of total histone H3 and H3 K9 acetylation revealed potential lower density of nucleosomes 2 kb upstream from the promoter region. Collectively, our data suggest that moderate level of alphaA-crystallin gene expression in lens epithelial cells does not require the presence of CBP and p300 in the promoter. However, the lens-specific chromatin domain contains both promoter localized CBP on the "background" of locus-spread presence of CBP and p300.