35 research outputs found
Primary and secondary structure of hamster vimentin predicted from the nucleotide sequence.
Tissue-specific activity of the blind mole rat and the two nucleotide-mutated mouse αB-crystallin promoter in transgenic mice
The αB-crystallin and HspB2 genes are located ≈0.9 kb apart in a head-to-head arrangement in mammals. Previous experiments have shown that a truncated −668/+45 αB-crystallin enhancer/promoter fragment from blind mole rats (Spalax ehrenbergi), which have nonfunctional lenses, lacks lens activity and has enhanced muscle activity in transgenic mice. Here we show that the full-length mole rat αB-crystallin intergenic region behaves similarly in transgenic mice. A two-nucleotide mutation ((−273)CA→G) in the mouse αB-crystallin enhancer/promoter fragment mimicking the wild-type mole rat sequence functionally converted the mouse promoter fragment to that of the wild-type mole rat promoter when tested in transgenic mice. The reciprocal mutation in the mole rat promoter fragment ((−272)G→CA) did not affect its activity. Oligonucleotides from the wild-type mouse and mole rat αB-crystallin promoter region under study formed distinct complexes with nuclear proteins from cultured cells. The mouse mutant sequence lost binding ability, whereas the mutated mole rat sequence gained the ability to form a complex similar in size to that of the wild-type mouse oligonucleotide. Our data support the idea that blind mole rats' αB-crystallin promoter activity was modified during the evolution of subterranean life and shows that tissue-specific promoter activity can be modulated by changing as few as two apparently neutral nucleotides in the mouse αB-crystallin enhancer region, implying the importance of the context of regulatory sequences for promoter activity
1H-NMR spectroscopy of bovine lens beta-crystallin. The role of the betaB2-crystallin C-terminal extension in aggregation
Adaptive evolution of small heat shock protein/ αB-crystallin promoter activity of the blind subterranean mole rat, Spalax ehrenbergi
Blind mole rats have degenerated subcutaneous eyes that are visually nonfunctional. In this investigation, we have compared the tissue specificity of the small heat shock protein (shsp)/αB-crystallin promoter of the mole rat superspecies, Spalax ehrenbergi, with that of the mouse. Earlier experiments showed that mouse shsp/αB-crystallin promoter/enhancer activity is high in the lens and moderate in the heart and skeletal muscle of transgenic mice. Here, we show in transgenic mouse experiments using the firefly luciferase reporter gene that, despite relatively few changes in sequence, the mole rat shsp/αB-crystallin promoter/enhancer has selectively lost lens activity after 13.5 days of embryogenesis (E13.5). The ratios of mole rat/mouse promoter activity were 0.01 for lens, 1.7 for heart, and 13.6 for skeletal muscle in 8-wk-old transgenic mice. Our data indicate that the shsp/αB-crystallin promoter/enhancer has undergone adaptive changes corresponding to the subterranean evolution of the blind mole rat. We speculate that selective pressures on metabolic economy may have contributed to these tissue-specific modifications of promoter/enhancer function during adaptation to life underground