绘制真核生物核糖体中可被甲硫基丙基-EDTA·Fe(II)和EDTA·Fe(II)接触到的RNA区域。

Mapping RNA regions in eukaryotic ribosomes that are accessible to methidiumpropyl-EDTA.Fe(II) and EDTA.Fe(II).

作者信息

Han H, Schepartz A, Pellegrini M, Dervan P B

机构信息

Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena 91125.

出版信息

Biochemistry. 1994 Aug 23;33(33):9831-44. doi: 10.1021/bi00199a004.

DOI:10.1021/bi00199a004

PMID:8060991

Abstract

Methidiumpropyl-EDTA.Fe(II) [MPE.Fe(II)] and EDTA.Fe(II) were used to investigate the structure of Drosophila melanogaster ribosomes. Cleavage reactions were performed on intact ribosomes in cell lysates in vitro and analyzed by primer extension with reverse transcriptase using oligodeoxynucleotide primers. Regions of 18S and 28S ribosomal RNAs (rRNAs) which are accessible to MPE.Fe(II) and EDTA.Fe(II) are located almost exclusively within expansion segments. The accessibility of these regions to cleavage indicates that they are likely exposed on the surface of eukaryotic ribosomes. These results provide information about the overall tertiary structure of rRNA in ribosomes.

摘要

用甲磺酸丙酯 - 乙二胺四乙酸铁（II）[MPE.Fe(II)] 和乙二胺四乙酸铁（II）来研究黑腹果蝇核糖体的结构。在体外细胞裂解液中的完整核糖体上进行切割反应，并使用寡脱氧核苷酸引物通过逆转录酶进行引物延伸分析。18S和28S核糖体RNA（rRNA）中可被MPE.Fe(II) 和EDTA.Fe(II) 作用的区域几乎完全位于扩展片段内。这些区域对切割的可及性表明它们可能暴露在真核核糖体表面。这些结果提供了有关核糖体中rRNA整体三级结构的信息。