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一种新型光交联CTP类似物的合成、表征及其在光亲和标记大肠杆菌和T7 RNA聚合酶中的应用。

Synthesis and characterization of a new photocrosslinking CTP analog and its use in photoaffinity labeling E. coli and T7 RNA polymerases.

作者信息

Hanna M M, Zhang Y, Reidling J C, Thomas M J, Jou J

机构信息

Department of Botany and Microbiology, University of Oklahoma, Norman 73019.

出版信息

Nucleic Acids Res. 1993 May 11;21(9):2073-9. doi: 10.1093/nar/21.9.2073.

DOI:10.1093/nar/21.9.2073
PMID:7684833
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC309467/
Abstract

A new photocrosslinking CTP analog that functioned as a substrate during transcription was synthesized and used to photoaffinity label E. coli and bacteriophage T7 RNA polymerases. This analog, 5-((4-azidophenacyl)thio) cytidine-5'-triphosphate (5-APAS-CTP) contains an aryl azide group approximately 10 A from the nucleotide base and specifically replaced CTP during synthesis of RNA by both polymerases. Analog was placed at the 3' end or internally within RNA. Both polymerases inefficiently incorporated two 5-APAS-CMP molecules sequentially, as was found for the related 5-APAS-UMP. Analog was placed at the 3' end of RNA in transcription complexes paused at the site of Q-modification of E. coli RNA polymerase, downstream of the lambda PR' promoter (+16), a pause that requires specific DNA sequences but no apparent RNA hairpin. Crosslinking was examined in the presence and absence of the NusA protein, which enhances the transcriptional pause at this site and is required for Q modification of the polymerase. Crosslinking of the 3' end of the RNA to NusA was not observed, consistent with our earlier results involving a NusA-enhanced pause site downstream from an RNA hairpin.

摘要

合成了一种新型光交联CTP类似物，它在转录过程中作为底物发挥作用，并用于对大肠杆菌和噬菌体T7 RNA聚合酶进行光亲和标记。这种类似物，5-((4-叠氮苯甲酰基)硫代)胞苷-5'-三磷酸(5-APAS-CTP)，在距核苷酸碱基约10埃处含有一个芳基叠氮基团，并且在两种聚合酶合成RNA的过程中特异性地取代了CTP。类似物被置于RNA的3'端或内部。两种聚合酶都低效地依次掺入两个5-APAS-CMP分子，正如在相关的5-APAS-UMP中所发现的那样。类似物被置于在大肠杆菌RNA聚合酶Q修饰位点（λPR'启动子下游(+16)处）暂停的转录复合物中RNA的3'端，该暂停需要特定的DNA序列，但不需要明显的RNA发夹结构。在有和没有NusA蛋白的情况下检查交联情况，NusA蛋白可增强该位点的转录暂停，并且是聚合酶Q修饰所必需的。未观察到RNA的3'端与NusA的交联，这与我们早期涉及RNA发夹下游NusA增强的暂停位点的结果一致。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/95ee/309467/21de28f6cda3/nar00058-0064-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/95ee/309467/eccb39400044/nar00058-0063-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/95ee/309467/21de28f6cda3/nar00058-0064-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/95ee/309467/eccb39400044/nar00058-0063-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/95ee/309467/21de28f6cda3/nar00058-0064-a.jpg

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Synthesis and characterization of a new photocrosslinking CTP analog and its use in photoaffinity labeling E. coli and T7 RNA polymerases.一种新型光交联CTP类似物的合成、表征及其在光亲和标记大肠杆菌和T7 RNA聚合酶中的应用。
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NusA contacts nascent RNA in Escherichia coli transcription complexes.NusA在大肠杆菌转录复合物中与新生RNA接触。
J Mol Biol. 1995 Apr 7;247(4):547-58. doi: 10.1006/jmbi.1994.0161.
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NusA changes the conformation of Escherichia coli RNA polymerase at the binding site for the 3' end of the nascent RNA.NusA改变了大肠杆菌RNA聚合酶在新生RNA 3'端结合位点处的构象。
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本文引用的文献

1
Effects of NusA protein on transcription termination in the tryptophan operon of Escherichia coli.NusA蛋白对大肠杆菌色氨酸操纵子转录终止的影响。
Cell. 1982 Jul;29(3):945-51. doi: 10.1016/0092-8674(82)90457-3.
2
RNA polymerase pausing and transcript release at the lambda tR1 terminator in vitro.体外λ噬菌体tR1终止子处的RNA聚合酶暂停与转录物释放
J Biol Chem. 1983 Aug 10;258(15):9391-7.
3
The phage lambda Q gene product: activity of a transcription antiterminator in vitro.噬菌体λQ基因产物：体外转录抗终止子的活性
利用一种新型光交联ATP类似物使RNA与RNA内部位置的AMP残基发生RNA-蛋白质交联。
Nucleic Acids Res. 2000 May 1;28(9):1849-58. doi: 10.1093/nar/28.9.1849.
4
Synthesis and evaluation of oligodeoxyribonucleotides containing an aryl(trifluoromethyl)diazirine moiety as the cross-linking probe: photoaffinity labeling of mammalian DNA polymerase beta.含芳基（三氟甲基）二氮丙啶部分作为交联探针的寡脱氧核糖核苷酸的合成与评价：哺乳动物DNA聚合酶β的光亲和标记
Nucleic Acids Res. 1997 Jun 15;25(12):2352-8. doi: 10.1093/nar/25.12.2352.
5
A photolabile 2',3'-dideoxyuridylate analog bearing an aryl(trifluoromethyl)diazirine moiety: photoaffinity labeling of HIV-1 reverse transcriptase.一种带有芳基（三氟甲基）二氮杂环丙烷部分的光不稳定2'，3'-二脱氧尿苷酸类似物：HIV-1逆转录酶的光亲和标记
Nucleic Acids Res. 1996 Sep 1;24(17):3364-9. doi: 10.1093/nar/24.17.3364.
6
NusA interferes with interactions between the nascent RNA and the C-terminal domain of the alpha subunit of RNA polymerase in Escherichia coli transcription complexes.在大肠杆菌转录复合物中，NusA干扰新生RNA与RNA聚合酶α亚基C末端结构域之间的相互作用。
Proc Natl Acad Sci U S A. 1995 May 23;92(11):5012-6. doi: 10.1073/pnas.92.11.5012.
7
Preparation of probe-modified RNA with 5-mercapto-UTP for analysis of protein-RNA interactions.用5-巯基尿苷三磷酸制备用于蛋白质-RNA相互作用分析的探针修饰RNA。
Nucleic Acids Res. 1995 Apr 11;23(7):1231-8. doi: 10.1093/nar/23.7.1231.
8
Site-specific RNA crosslinking with 4-thiouridine.使用4-硫尿苷进行位点特异性RNA交联。
Mol Biol Rep. 1994 Jul;20(1):35-44. doi: 10.1007/BF00999853.
9
NusA changes the conformation of Escherichia coli RNA polymerase at the binding site for the 3' end of the nascent RNA.NusA改变了大肠杆菌RNA聚合酶在新生RNA 3'端结合位点处的构象。
J Bacteriol. 1994 Mar;176(6):1787-9. doi: 10.1128/jb.176.6.1787-1789.1994.
Cell. 1982 Sep;30(2):637-48. doi: 10.1016/0092-8674(82)90260-4.
4
Toxicity of an overproduced foreign gene product in Escherichia coli and its use in plasmid vectors for the selection of transcription terminators.大肠杆菌中外源基因过量表达产物的毒性及其在用于转录终止子选择的质粒载体中的应用。
Gene. 1984 Feb;27(2):161-72. doi: 10.1016/0378-1119(84)90137-9.
5
Pausing and attenuation of in vitro transcription in the rrnB operon of E. coli.大肠杆菌rrnB操纵子中体外转录的暂停和衰减
Cell. 1981 Dec;27(3 Pt 2):523-31. doi: 10.1016/0092-8674(81)90394-9.
6
Nucleosides. XXXVII. 5,6-substituted 5-fluorodihydropyrimidines and their 2'-deoxyribonucleosides.核苷。XXXVII。5,6-取代的5-氟二氢嘧啶及其2'-脱氧核糖核苷。
J Med Chem. 1967 Jan;10(1):47-58. doi: 10.1021/jm00313a010.
7
Phage lambda gene Q antiterminator recognizes RNA polymerase near the promoter and accelerates it through a pause site.噬菌体λ基因Q抗终止子在启动子附近识别RNA聚合酶，并使其加速通过一个暂停位点。
Cell. 1985 Aug;42(1):259-69. doi: 10.1016/s0092-8674(85)80121-5.
8
nusA protein of Escherichia coli is an efficient transcription termination factor for certain terminator sites.大肠杆菌的nusA蛋白是某些终止子位点的高效转录终止因子。
J Mol Biol. 1987 Jun 20;195(4):809-18. doi: 10.1016/0022-2836(87)90486-4.
9
Gene Q antiterminator proteins of Escherichia coli phages 82 and lambda suppress pausing by RNA polymerase at a rho-dependent terminator and at other sites.大肠杆菌噬菌体82和λ的基因Q抗终止蛋白可抑制RNA聚合酶在ρ因子依赖性终止子及其他位点处的暂停。
Proc Natl Acad Sci U S A. 1989 Jul;86(14):5301-5. doi: 10.1073/pnas.86.14.5301.
10
Photoaffinity cross-linking methods for studying RNA-protein interactions.用于研究RNA-蛋白质相互作用的光亲和交联方法。
Methods Enzymol. 1989;180:383-409. doi: 10.1016/0076-6879(89)80113-2.