GENERATION OF HISTONE MESSENGER-RNA 3' ENDS BY ENDONUCLEOLYTIC CLEAVAGE OF THE PRE-MESSENGER-RNA IN A SNRNP-DEPENDENT INVITRO REACTION
GENERATION OF HISTONE MESSENGER-RNA 3' ENDS BY ENDONUCLEOLYTIC CLEAVAGE OF THE PRE-MESSENGER-RNA IN A SNRNP-DEPENDENT INVITRO REACTION
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DOI：
10.1002/j.1460-2075.1986.tb04362.x
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发表时间：
1986-06-01
期刊：
影响因子：
11.4
通讯作者：
BIRNSTIEL, ML
中科院分区：
文献类型：
--
作者：
GICK, O;KRAMER, A;BIRNSTIEL, ML
SP 6产生的小鼠组蛋白H4 mRNA前体在HeLa细胞的核提取物中孵育，产生经过加工的mRNA物质，其末端位于保守末端ACCA序列的3 ''腺苷上，与从序列中分离的十种不同的组蛋白mRNA不同，与从海胆胚胎中分离的十种不同的组蛋白mRNA不同，其末端位于3 '' C或A。在存在20 mM EDTA的情况下，RNA转录物的3“间隔区部分被切割掉，因此组蛋白mRNA的3”末端是前体RNA的核酸内切切割的结果。在体外裂解反应是特异性抑制的Sm-血清型的人抗血清，但不是由对照血清，可以通过添加制备的部分纯化的小核RNP的抗体耗尽的提取物获救。有趣的是，snRNP制剂足以在不存在添加的核提取物的情况下引发前mRNA的3“加工。
Incubation of SP6 generated mouse histone H4 mRNA precursors in nuclear extracts of HeLa cells yields processed mRNA species which end on the 3'' adenosine of the conserved terminal ACCA sequence not unlike ten different histone mRNAs isolated from sequence not unlike ten different histone mRNAs isolated from sea urchin embryos which end either on the 3'' C or A. In the presence of 20 mM EDTA, the cleaved off 3'' spacer portions of the RNA transcripts are readily detected, hence the 3'' ends of histone mRNAs arise as a consequence of endonucleolytic cleavage(s) of the precursor RNA. The in vitro cleavage reaction is specifically inhibited by human antisera of the Sm-serotype, but not by control sera and can be rescued by the addition of a preparation of partially purified small nuclear RNPs to the antibody-depleted extract. Interestingly, the snRNP preparation is sufficient to elicit 3'' processing of pre-mRNA in the absence of added nuclear extract.