通过补骨脂素交联确定的M1 RNA结构。

Structure of M1 RNA as determined by psoralen cross-linking.

作者信息

Lipson S E, Cimino G D, Hearst J E

机构信息

Department of Chemistry, University of California, Berkeley 94720.

出版信息

Biochemistry. 1988 Jan 26;27(2):570-5. doi: 10.1021/bi00402a011.

DOI:10.1021/bi00402a011

PMID:2450574

Abstract

The RNA moiety of ribonuclease P from Escherichia coli (M1 RNA) has been photoreacted with 4'-(hydroxymethyl)-4,5',8-trimethylpsoralen (HMT) and long-wave UV light (320-380 nm) in a buffer containing 60 mM Mg2+, where the RNA moiety acts as a true catalyst of tRNA processing. Limited specific digestion and two-dimensional gel electrophoresis yield fragments cross-linked by HMT. By photoreversal of the isolated cross-linked fragments and enzymatic sequencing of the fragments, the positions of the cross-links have been elucidated. This method allows us to locate the cross-link to +/- 15 nucleotides. Further assignments of the exact locations of the cross-links have been made on the basis of the known photoreactivity of the psoralen with different bases. Nine unique cross-links have been isolated in the M1 RNA including four long-range interactions. The short-range interactions are discussed here in detail.

摘要

来自大肠杆菌的核糖核酸酶P的RNA部分（M1 RNA）已在含有60 mM Mg2+的缓冲液中，与4'-（羟甲基）-4,5',8-三甲基补骨脂素（HMT）和长波紫外线（320 - 380 nm）进行光反应，在此RNA部分作为tRNA加工的真正催化剂。有限的特异性消化和二维凝胶电泳产生了由HMT交联的片段。通过对分离的交联片段进行光逆转和对片段进行酶促测序，已阐明交联的位置。这种方法使我们能够将交联定位到±15个核苷酸。基于补骨脂素与不同碱基的已知光反应性，对交联的确切位置进行了进一步的确定。在M1 RNA中已分离出九个独特的交联，包括四个长程相互作用。本文将详细讨论短程相互作用。