ARD-1 cDNA from human cells encodes a site-specific single-strand endoribonuclease that functionally resembles Escherichia coli RNase E
ARD-1 cDNA from human cells encodes a site-specific single-strand endoribonuclease that functionally resembles Escherichia coli RNase E
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DOI：
10.1074/jbc.272.21.13823
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发表时间：
1997-05-23
影响因子：
4.8
通讯作者：
Cohen, SN
中科院分区：
文献类型：
--
作者：
ClaverieMartin, F;Wang, M;Cohen, SN
人ARD-I（RNA衰变激活剂）cDNA序列可以拯救大肠杆菌rne基因中的突变，该基因特异性地产生必需的核糖核酸内切酶RNase E，导致在体内在me缺陷型细菌中以及在体外在从这些细胞分离的提取物中产生RNase E样裂解（Wang，M.，和Cohen，S. N.等人（1994）Proc. Acad. Sci.联合S. A. 91，10591-10595）。最近的研究表明，由ARD-I cDNA编码的13.3-kDa蛋白几乎与牛蛋白NIPP-1（蛋白磷酸酶1的核抑制剂）的羧基末端相同;提出通过可变剪接形成的单独转录物编码离散的ARD-1和组合的ARD-1/NIPP-1产物（货车Eynde，A.，Wera，S.，Beullens，M.，Torrekens，S.，货车鲁汶，F.，Stalmans，W.，和Bollens，M.（1995）J.Biol.Chem.270，28068-28074）。在这里，我们表明亲和柱纯化的蛋白编码的人ARD-I cDNA在E。大肠杆菌中的核糖核酸内切酶是一种位点特异性的Mg 2+依赖性核糖核酸内切酶，其在体外与RNase E底物结合，在与RNase E相同的位点切割RNA，并且像RNase E一样，在切割位点产生5'磷酸末端。我们的研究结果表明，ARD-I肽可以作为一个核糖核酸类似物关闭。coli RNase E以及蛋白磷酸酶抑制剂NIPP-1的结构域。
The human ARD-I (activator of RNA decay) cDNA sequence can rescue mutations in the Escherichia coli rne gene, which specifies the essential endoribonuclease RNase E, resulting in RNase E-like cleavages in vivo in me-defective bacteria and in vitro in extracts isolated from these cells (Wang, M., and Cohen, S. N. (1994) Proc. Natl. Acad. Sci. U. S. A. 91, 10591-10595). Recent studies indicate that the 13.3-kDa protein encoded by ARD-I cDNA is almost identical to the carboxyl-terminal end of the bovine protein NIPP-1, a nuclear inhibitor of protein phosphatase 1; separate transcripts formed by alternative splicing are proposed to encode the discrete ARD-1 and combined ARD-1/NIPP-1 products (Van Eynde, A., Wera, S., Beullens, M., Torrekens, S., Van Leuven, F., Stalmans, W., and Bollens, M. (1995) J. Biol. Chem. 270, 28068-28074). Here we show that affinity column-purified protein encoded by human ARD-I cDNA in E. coli is a site-specific Mg2+-dependent endoribonuclease that binds in vitro to RNase E substrates, cleaves RNA at the same sites as RNase E, and, like RNase E, generates 5' phosphate termini at sites of cleavage. Our results indicate that the ARD-I peptide can function as a ribonucleolytic analog off. coli RNase E as well as a domain of the protein phosphatase inhibitor, NIPP-1.