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In vitro 3' end processing and poly(A) tailing of RNA in Trypanosoma cruzi.

作者信息

Zwierzynski T A, Widmer G, Buck G A

机构信息

Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond 23298.

出版信息

Nucleic Acids Res. 1989 Jun 26;17(12):4647-60. doi: 10.1093/nar/17.12.4647.

DOI:10.1093/nar/17.12.4647
PMID:2473439
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC318022/
Abstract

Pre-mRNA in kinetoplastids is processed to maturity following unique pathways requiring a transplicing event that links a common 39 nucleotide leader to the 5' termini of the mature mRNAs. The mechanisms of this reaction and other steps of mRNA processing; i.e., 5' capping and 3' cleavage and polyadenylation, have not been resolved. Herein, we describe a 3' polyadenylation activity in cell-free extracts prepared from nuclei isolated from Trypanosoma cruzi, the kinetoplastid agent of Chagas' Disease. Synthetic RNA transcripts incubated in these extracts in the presence of ATP are 3' polyadenylated. This polyadenylation activity is sensitive to heat or pre-treatment of the extract with Micrococcal nuclease, suggesting that an RNA-protein complex is required. As these are characteristics of polyadenylation activities in other eukaryotes, we believe that this activity may participate in the in vivo trypanosome mRNA polyadenylation system. Several other modification activities specific for RNA 3' termini, including terminal nucleotide transferases, a tRNA CCA maturation activity, and a 3' exonuclease were also identified in these T. cruzi nuclear extracts.

摘要
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a4a/318022/f0396e5c8660/nar00129-0233-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a4a/318022/a3d5a2394ca1/nar00129-0226-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a4a/318022/8426e3904841/nar00129-0227-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a4a/318022/874d31e57e1f/nar00129-0229-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a4a/318022/53345643386d/nar00129-0230-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a4a/318022/32170a1c1018/nar00129-0231-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a4a/318022/f0396e5c8660/nar00129-0233-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a4a/318022/a3d5a2394ca1/nar00129-0226-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a4a/318022/8426e3904841/nar00129-0227-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a4a/318022/874d31e57e1f/nar00129-0229-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a4a/318022/53345643386d/nar00129-0230-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a4a/318022/32170a1c1018/nar00129-0231-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a4a/318022/f0396e5c8660/nar00129-0233-a.jpg

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引用本文的文献

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Transcription termination and 3'-End processing of the spliced leader RNA in kinetoplastids.动质体中剪接前导RNA的转录终止及3'末端加工
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Molecular cloning and characterization of the 78-kilodalton glucose-regulated protein of Trypanosoma cruzi.

本文引用的文献

1
Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.从分离的哺乳动物细胞核的可溶性提取物中，RNA聚合酶II进行准确的转录起始。
Nucleic Acids Res. 1983 Mar 11;11(5):1475-89. doi: 10.1093/nar/11.5.1475.
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Recognition of cap structure in splicing in vitro of mRNA precursors.体外mRNA前体剪接中帽结构的识别
Cell. 1984 Oct;38(3):731-6. doi: 10.1016/0092-8674(84)90268-x.
3
Strains and clones of Trypanosoma cruzi can be characterized by pattern of restriction endonuclease products of kinetoplast DNA minicircles.
克氏锥虫78千道尔顿葡萄糖调节蛋白的分子克隆与特性分析
Infect Immun. 1994 Jun;62(6):2499-507. doi: 10.1128/iai.62.6.2499-2507.1994.
4
In vitro capping in Trypanosoma cruzi identifies and shows specificity for the spliced leader RNA and U-RNAs.克氏锥虫中的体外加帽作用鉴定出剪接前导RNA和U-RNA并显示出特异性。
Nucleic Acids Res. 1990 Jul 25;18(14):4197-206. doi: 10.1093/nar/18.14.4197.
5
RNA-protein complexes mediate in vitro capping of the spliced-leader primary transcript and U-RNAs in Trypanosoma cruzi.RNA-蛋白质复合物介导克氏锥虫中剪接前导序列初级转录本和U-RNA的体外加帽。
Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5626-30. doi: 10.1073/pnas.88.13.5626.
6
RNA editing in trypanosomes. The us(e) of guide RNAs.锥虫中的RNA编辑。向导RNA的作用。
Mol Biol Rep. 1992 Sep;16(4):217-27. doi: 10.1007/BF00419661.
克氏锥虫的菌株和克隆可通过动基体DNA微小环的限制性内切酶产物模式来进行表征。
Proc Natl Acad Sci U S A. 1980 Nov;77(11):6810-4. doi: 10.1073/pnas.77.11.6810.
4
Splicing of in vitro synthesized messenger RNA precursors in HeLa cell extracts.体外合成的信使核糖核酸前体在HeLa细胞提取物中的剪接
Cell. 1983 Nov;35(1):89-99. doi: 10.1016/0092-8674(83)90211-8.
5
Specific labeling of 3' termini of RNA with T4 RNA ligase.利用T4 RNA连接酶对RNA的3'末端进行特异性标记。
Methods Enzymol. 1980;65(1):65-74. doi: 10.1016/s0076-6879(80)65011-3.
6
Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter.从含有噬菌体SP6启动子的质粒中高效体外合成生物活性RNA和RNA杂交探针。
Nucleic Acids Res. 1984 Sep 25;12(18):7035-56. doi: 10.1093/nar/12.18.7035.
7
Electrophoretic separation of polyadenylation-specific complexes.多聚腺苷酸化特异性复合物的电泳分离
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Multiple factors are required for poly(A) addition to a mRNA 3' end.在mRNA 3'末端添加多聚腺苷酸（poly(A)）需要多种因素。
Genes Dev. 1988 May;2(5):588-97. doi: 10.1101/gad.2.5.588.
9
Cleavage and polyadenylation of messenger RNA precursors in vitro occurs within large and specific 3' processing complexes.信使核糖核酸前体在体外的切割和聚腺苷酸化发生在大型且特定的3'加工复合物内。
EMBO J. 1987 Dec 20;6(13):4159-68. doi: 10.1002/j.1460-2075.1987.tb02762.x.
10
Major transcript of the frameshifted coxII gene from trypanosome mitochondria contains four nucleotides that are not encoded in the DNA.来自锥虫线粒体的移码coxII基因的主要转录本包含四个未在DNA中编码的核苷酸。
Cell. 1986 Sep 12;46(6):819-26. doi: 10.1016/0092-8674(86)90063-2.