Cloning of a gene, YT521, for a novel RNA splicing-related protein induced by hypoxia/reoxygenation
Cloning of a gene, YT521, for a novel RNA splicing-related protein induced by hypoxia/reoxygenation
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DOI：
10.1016/s0169-328x(97)00262-3
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发表时间：
1998-01-01
期刊：
影响因子：
--
通讯作者：
Takagi, T
中科院分区：
文献类型：
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作者：
Imai, Y;Matsuo, N;Takagi, T
为了阐明星形胶质细胞在中枢神经系统对缺血的应激反应中的作用，在大鼠培养的星形胶质细胞暴露于缺氧/复氧后，早期基因表达进行了检查，我们以前克隆了一种新的RNA结合蛋白，RA 301，从复氧的星形胶质细胞。此外，我们现在已经克隆了一个新的基因RA 301结合蛋白，命名为YT 521，通过酵母双杂交筛选技术，探索RA 301的功能。YT 521 cDNA全长约3200 bp，开放阅读框编码712个氨基酸。该氨基酸序列含有精氨酸-天冬氨酸-谷氨酸富集区和谷氨酸富集区，并且与RNA结合蛋白如U1- 70 k具有低程度的同源性。北方印迹分析显示，缺氧后星形胶质细胞YT 521 mRNA表达上调。YT 521 mRNA的诱导是由内源性产生的活性氧介导的，因为它是通过用二苯基碘鎓处理细胞来抑制的，二苯基碘鎓阻断星形胶质细胞形成氧自由基。这些表达模式类似于RA 301 mRNA的表达模式。远距离Western blot分析表明，YT 521蛋白不仅与RA 301蛋白相互作用，还与剪接因子SC 35和SF 2相互作用。这些结果表明，YT 521是一个新的候选RNA剪接相关蛋白。(C)1998年Elsevier Science B.V.
To elucidate the role of astrocytes in the stress response of the central nervous system to ischemia, early gene expression was examined in rat cultured astrocytes after the exposure to hypoxia/reoxygenation, and we have previously cloned a novel RNA binding protein, RA301, from the reoxygenated astrocytes. Furthermore, we have now cloned a new gene for RA301 binding protein, termed YT521, by a yeast two-hybrid screening technique to explore RA301 functions. The YT521 cDNA is about 3200 bp long with an open reading frame encoding 712 amino acids. This amino acid sequence contains arginine-aspartic acid-glutamic acid rich region and glutamic acid rich one, and has a low degree of homology with RNA binding proteins such as U1-70k. Northern blot analysis revealed that YT521 mRNA expression was up-regulated in reoxygenated astrocytes. Induction of YT521 mRNA was mediated by endogenously generated reactive oxygen species, as it was suppressed by treatment of the cells with diphenyl iodonium which blocks oxygen-free radical formation by astrocytes. These expression patterns resembled those of RA301 mRNA. Far Western blot analysis showed that YT521 protein was not only interacting with RA301 protein, but also with SC35 and SF2, both of which are splicing factors. These results suggest that YT521 is a novel candidate for RNA splicing-related protein. (C) 1998 Elsevier Science B.V.