UV-crosslinking of E1 small nucleolar RNA to proteins in frog oocytes.
UV-crosslinking of E1 small nucleolar RNA to proteins in frog oocytes.
复制标题
E1 小核仁 RNA 与青蛙卵母细胞中蛋白质的紫外交联。
DOI：
10.1002/jcp.20223
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发表时间：
2005
期刊：
影响因子：
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通讯作者：
Eliceiri,GeorgeL
中科院分区：
文献类型：
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作者：
Smith,JamesL;Walton,AndrewH;Eliceiri,GeorgeL
E1/U17小核仁RNA（small nucleolar RNA，snoRNA）是一种盒H/ACA snoRNA。为了检测主要在尿苷处与E1 RNA发生UV交联的蛋白条带，将[α-32 P]UTP标记的E1 RNA注入蛙卵母细胞并孵育，对分离的细胞核进行UV照射，并用RNase A消化细胞核内容物。野生型E1 RNA特异性UV交联至几个蛋白条带。为了确定参与这些相互作用的E1 RNA位点，我们测试了21个E1 RNA突变体，每个突变体由保守序列或结构中的取代组成。不同蛋白质条带与E1 RNA的UV交联依赖于以下保守的E1 RNA片段之一：两个5′端RNA位点;五个5′半RNA位点;两个3′半RNA位点;或位于整个E1 RNA中的14个位点。在这些保守的E1 RNA位点中，UV交联显然依赖于11个位点的序列和2个位点的结构。有和没有RNA竞争的凝胶电泳检测到的蛋白质条带是不常见的所有盒H/ACA snoRNA。© 2004 Wiley利斯公司
E1/U17 small nucleolar RNA (snoRNA) is a box H/ACA snoRNA. To detect protein bands that UV‐crosslink to E1 RNA primarily at uridines, frog oocytes were injected with [α‐32P]UTP‐labeled E1 RNA and incubated, isolated nuclei were UV irradiated, and nuclear contents were digested with RNase A. Wild‐type E1 RNA specifically UV‐crosslinked to several protein bands. To identify E1 RNA sites involved in these interactions, we tested 21 E1 RNA mutants, each consisting of substitutions in a conserved sequence or structure. UV‐crosslinking of different protein bands to E1 RNA depended on one of the following sets of conserved E1 RNA segments: two 5′ end RNA sites; five 5′ half RNA sites; two 3′ half RNA sites; or 14 sites located throughout E1 RNA. Of these conserved E1 RNA sites, UV‐crosslinking apparently depended on sequences at 11 sites, and structures at 2 sites. Gel electrophoresis with and without RNA competition detected protein bands that are not common to all of the box H/ACA snoRNAs. © 2004 Wiley‐Liss, Inc.