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Mark R. Verardo, Andrea Viczian, Natik Piri, Novrouz B. Akhmedov, Barry E. Knox, Debora B. Farber; Regulatory Sequences in the 3′ Untranslated Region of the Human cGMP-Phosphodiesterase β-Subunit Gene. Invest. Ophthalmol. Vis. Sci. 2009;50(6):2591-2598. doi: 10.1167/iovs.08-2010.
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purpose. Rod cGMP-phosphodiesterase, a key enzyme in visual transduction, is important for retinal integrity and function. Mutations in the gene encoding the phosphodiesterase β-subunit (PDEβ) cause retinal degeneration in animals and humans. Here the authors tested the hypothesis that elements in the 3′ untranslated region (3′ UTR) of the PDEβ gene are involved in the regulation of PDEβ expression.
methods. Involvement of the 3′ UTR of PDEβ mRNA in the regulation of PDEβ expression was assessed by Y-79 retinoblastoma cells or the heads of Xenopus laevis tadpoles with constructs containing the SV40 or PDEβ promoter, the luciferase cDNA, and either the SV40 or the PDEβ 3′ UTR (or fragments of its sequence).
results. Compared with the SV40 3′ UTR (used as control), the entire PDEβ 3′ UTR decreased reporter gene expression in Y-79 retinoblastoma cells as well as in SY5Y neuroblastoma and 293 human embryonic kidney cell lines. However, the authors observed that two 100-nucleotide fragments from the PDEβ 3′ UTR increased while its noncanonical poly-adenylation signal abolished reporter gene expression in Y-79 retinoblastoma cells and in ex vivo experiments using Xenopus tadpole heads. In particular, an 11-nucleotide element (EURE) in one of the 100-nucleotide fragments was responsible for the upregulation of luciferase expression.
conclusions. These studies indicate that the 3′ UTR of the PDEβ mRNA is involved in the complex regulation of this gene’s expression in the retina. Moreover, the results show that the PDEβ poly-A signal has a dominant inhibitory effect over two other regions in the 3′ UTR that stimulate gene expression.
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