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酵母tRNALeu3基因的突变前体中的结构改变，这些突变前体对于一种高度纯化的剪接内切核糖核酸酶而言表现为有缺陷的底物。

Structural alterations in mutant precursors of the yeast tRNALeu3 gene which behave as defective substrates for a highly purified splicing endoribonuclease.

作者信息

Attardi D G, Margarit I, Tocchini-Valentini G P

出版信息

EMBO J. 1985 Dec 1;4(12):3289-97. doi: 10.1002/j.1460-2075.1985.tb04079.x.

DOI:10.1002/j.1460-2075.1985.tb04079.x
PMID:3937725
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC554656/
Abstract

We have produced a highly purified preparation of the Xenopus laevis splicing endonuclease (XlaI RNase). The purified enzyme correctly cleaves tRNA precursors, creating substrates for subsequent ligation. The 5'-half molecules have a 2',3' cyclic phosphate at their 3' termini. Assuming that splicing enzymes recognize primarily structural elements in the 'mature domain', we have been studying the conformation of three splicing-defective precursors made from mutants of the yeast tRNALeu3 gene. The mutations alter base-pairing in the D-stem region and two of the mutants are absolute defectives. Enzymatic probing of the structures of the altered tRNA precursors shows that the structural perturbations in these mutants are localized on the 'inside' of the 'L'-shaped three-dimensional structure. The implications of this finding for the recognition process are discussed.

摘要

我们制备了一种高度纯化的非洲爪蟾剪接内切核酸酶（XlaI核糖核酸酶）制剂。纯化后的酶能正确切割tRNA前体，为后续连接反应生成底物。5'半分子在其3'末端具有2'，3'环磷酸酯。假设剪接酶主要识别“成熟结构域”中的结构元件，我们一直在研究由酵母tRNALeu3基因突变体产生的三种剪接缺陷前体的构象。这些突变改变了D茎区域的碱基配对，其中两个突变体是绝对缺陷型。对改变后的tRNA前体结构进行酶促探测表明，这些突变体中的结构扰动位于“L”形三维结构的“内部”。讨论了这一发现对识别过程的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb3b/554656/882fcfe84d6c/emboj00277-0248-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb3b/554656/a462705f0282/emboj00277-0244-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb3b/554656/7b115aad6081/emboj00277-0245-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb3b/554656/1630f7b6d26a/emboj00277-0245-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb3b/554656/bba48a5239a8/emboj00277-0246-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb3b/554656/edd63674a3b3/emboj00277-0247-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb3b/554656/882fcfe84d6c/emboj00277-0248-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb3b/554656/a462705f0282/emboj00277-0244-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb3b/554656/7b115aad6081/emboj00277-0245-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb3b/554656/1630f7b6d26a/emboj00277-0245-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb3b/554656/bba48a5239a8/emboj00277-0246-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb3b/554656/edd63674a3b3/emboj00277-0247-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb3b/554656/882fcfe84d6c/emboj00277-0248-a.jpg

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本文引用的文献

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A covalent adduct between the uracil ring and the active site of an aminoacyl tRNA synthetase.尿嘧啶环与氨酰tRNA合成酶活性位点之间的共价加合物。
Nature. 1982 Jul 8;298(5870):136-40. doi: 10.1038/298136a0.
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Transcription and processing of a yeast tRNA gene containing a modified intervening sequence.一个含有修饰性间隔序列的酵母tRNA基因的转录与加工
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Junction phosphate is derived from the precursor in the tRNA spliced by the archaeon Haloferax volcanii cell extract.连接磷酸是由嗜盐嗜碱古菌沃氏嗜盐菌细胞提取物剪接的tRNA前体衍生而来。
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The human RNA 3'-terminal phosphate cyclase is a member of a new family of proteins conserved in Eucarya, Bacteria and Archaea.人类RNA 3'-末端磷酸环化酶是真核生物、细菌和古细菌中保守的一个新蛋白质家族的成员。
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Substrate recognition and identification of splice sites by the tRNA-splicing endonuclease and ligase from Saccharomyces cerevisiae.酿酒酵母中tRNA剪接内切核酸酶和连接酶对剪接位点的底物识别与鉴定
Mol Cell Biol. 1987 Jan;7(1):76-84. doi: 10.1128/mcb.7.1.76-84.1987.
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Pre-tRNA splicing in a nuclear extract from human leukaemia cells: separation of endonuclease and ligase activities.人白血病细胞核提取物中的前体tRNA剪接：核酸内切酶和连接酶活性的分离
Nucleic Acids Res. 1988 Aug 11;16(15):7713-4. doi: 10.1093/nar/16.15.7713.
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