...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Molecular and Cellular Biology >Expression and regulation by interferon of a double-stranded-RNA-specific adenosine deaminase from human cells: evidence for two forms of the deaminase.
【24h】

Expression and regulation by interferon of a double-stranded-RNA-specific adenosine deaminase from human cells: evidence for two forms of the deaminase.

机译：干扰素从人细胞表达和调节双链RNA特异性腺苷脱氨酶：两种形式的脱氨酶的证据。

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

A 6,474-nucleotide human cDNA clone designated K88, which encodes double-stranded RNA (dsRNA)-specific adenosine deaminase, was isolated in a screen for interferon (IFN)-regulated cDNAs. Northern (RNA) blot analysis revealed that the K88 cDNA hybridized to a single major transcript of approximately 6.7 kb in human cells which was increased about fivefold by IFN treatment. Polyclonal antisera prepared against K88 cDNA products expressed in Escherichia coli as glutathione S-transferase (GST) fusion proteins recognized two proteins by Western (immunoblot) analysis. An IFN-induced 150-kDa protein and a constitutively expressed 110-kDa protein whose level was not altered by IFN treatment were detected in human amnion U and neuroblastoma SH-SY5Y cell lines. Only the 150-kDa protein was detected in mouse fibroblasts with antiserum raised against the recombinant human protein; the mouse 150-kDa protein was IFN inducible. Immunofluorescence microscopy and cell fractionation analyses showed that the 110-kDa protein was exclusively nuclear, whereas the 150-kDa protein was present in both the cytoplasm and nucleus of human cells. The amino acid sequence deduced from the K88 cDNA includes three copies of the highly conserved R motif commonly found in dsRNA-binding proteins. Both the 150-kDa and the 110-kDa proteins prepared from human nuclear extracts bound to double-stranded but not to single-stranded RNA affinity columns. Furthermore, E. coli-expressed GST-K88 fusion proteins that included the R motif possessed dsRNA-binding activity. Extracts prepared either from K88 cDNA-transfected cells or from IFN-treated cells contained increased dsRNA-specific adenosine deaminase enzyme activity. These results establish that K88 encodes an IFN-inducible dsRNA-specific adenosine deaminase and suggest that at least two forms of dsRNA-specific adenosine deaminase occur in human cells.
机译：在筛选干扰素（IFN）调控的cDNA的过程中，分离出了一个称为K88的6,474个核苷酸的人类cDNA克隆，该克隆编码双链RNA（dsRNA）特异性腺苷脱氨酶。 Northern（RNA）印迹分析表明，K88 cDNA与人细胞中约6.7 kb的单个主要转录物杂交，通过IFN处理，该转录物增加了约5倍。针对在大肠杆菌中以谷胱甘肽S-转移酶（GST）融合蛋白表达的K88 cDNA产物制备的多克隆抗血清通过Western（免疫印迹）分析识别了两种蛋白。在人羊膜U和神经母细胞瘤SH-SY5Y细胞系中检测到IFN诱导的150 kDa蛋白和组成型表达的110 kDa蛋白，其水平未通过IFN处理改变。在小鼠成纤维细胞中仅检测到150kDa的蛋白质，并且产生了针对重组人蛋白质的抗血清。小鼠150 kDa蛋白可被IFN诱导。免疫荧光显微镜和细胞分级分析表明，110 kDa蛋白仅是核蛋白，而150 kDa蛋白则存在于人细胞的细胞质和细胞核中。从K88 cDNA推导的氨基酸序列包括三个副本的dsRNA结合蛋白中常见的高度保守的R基序。从人核提取物制备的150 kDa和110 kDa蛋白都与双链而不是单链RNA亲和柱结合。此外，包括R基序的大肠杆菌表达的GST-K88融合蛋白具有dsRNA结合活性。从K88 cDNA转染的细胞或经IFN处理的细胞制备的提取物含有增加的dsRNA特异性腺苷脱氨酶活性。这些结果证实，K88编码IFN诱导的dsRNA特异性腺苷脱氨酶，并且表明在人细胞中至少存在两种​​形式的dsRNA特异性腺苷脱氨酶。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号