Identification of 23S rRNA nucleotides neighboring the P-loop in the Escherichia coli 50S subunit.

Identification of 23S rRNA nucleotides neighboring the P-loop in the Escherichia coli 50S subunit.
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鉴定大肠杆菌 50S 亚基中 P 环附近的 23S rRNA 核苷酸。

DOI:
10.1093/nar/27.22.4376
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发表时间:
1999
影响因子:
14.9
通讯作者:
Cooperman,BS
Cooperman,BS
中科院分区:
生物学2区
文献类型:
--
作者:
Bukhtiyarov,Y;Druzina,Z;Cooperman,BS

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本文报道了一种放射性、光不稳定的2′-O-甲基寡RNA探针2258-53/52(SAz)-48,PHONT 1的合成及其在鉴定23 SrRNA P环附近核苷酸中的应用。该探针与大肠杆菌50 S亚基的nt 2248-2258互补。PHONT 1含有连接到与位置2252-2253互补的核苷酸之间的硫代磷酸酯桥上的对叠氮苯酰基,使得光生氮宾最大距离23 S RNA核苷酸G2252和G2253 17-19个碱基。PHONT 1与50 S亚基结合,并在其靶位点内或紧邻其靶位点处光掺入,以及掺入落在G2357和A2430之间的几个核苷酸中。这些结果的意义的肽基转移酶中心的结构被认为是。PHONT方法通常适用于复杂的含RNA分子的研究。
We report the synthesis of a radioactive, photolabile 2′-O-methyloligoRNA probe, 2258-53/52(SAz)-48, PHONT1, and its exploitation in identifying 23S rRNA nucleotides neighboring the so-called ′P-loop′. The probe is complementary to nt 2248-2258 inEscherichia coli50S subunits. PHONT 1 contains a p-azidophenacyl group attached to a phosphorothioate bridge between the nucleotides complementary to the positions 2252–2253, such that the photogenerated nitrene is maximally 17-19 Å from 23S RNA nucleotides G2252 and G2253. PHONT 1 binds to the 50S subunit, and photoincorporates within or immediately adjacent to its target site, as well as into several nucleotides falling between G2357 and A2430. The significance of these results for the structure of the peptidyl trans-ferase center is considered. The PHONT approach is generally applicable to studies of complex RNA-containing molecules.
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