硫醇化氨酰基转移核糖核酸与核糖体核糖核酸及蛋白质的光交联作用。

Photocrosslinking of thiolated aminoacyl-tRNA to ribosomal RNA and proteins.

作者信息

Barritault D, Buckingham R H, Favre A, Thomas G

出版信息

Biochimie. 1981 Jul;63(7):587-93. doi: 10.1016/s0300-9084(81)80056-9.

Abstract

tRNA has been converted to a form that can be photoactivated by chemical modification of some of the exposed cytidine residues to thio-4-uridine A certain percentage of the modified molecules can be charged and bound to the ribosome; thiolated fMet-tRNAfMet is bound to the P-site as shown by puromycin reactivity. Near the UV irradiation produces covalent crosslinks between total thiolated AA-tRNA or fMet-tRNAfMet and the ribosome. AA-tRNA becomes crosslinked to both 30S and 50S subunits but fMet-tRNAfMet to 50S subunits alone. In each case, crosslinking of tRNA was found to be not only to ribosomal proteins, but also to rRNA. The covalent complexes appear sufficiently stable to allow identification of the proteins or rRNA sequences involved.

摘要

转运RNA（tRNA）已被转化为一种可通过将一些暴露的胞嘧啶残基化学修饰为硫代-4-尿苷而进行光活化的形式。一定比例的修饰分子可以被装载并与核糖体结合；如嘌呤霉素反应所示，硫醇化的甲硫氨酸起始tRNA（fMet-tRNAfMet）结合到P位点。紫外线照射会使总的硫醇化氨基酸-tRNA或fMet-tRNAfMet与核糖体之间产生共价交联。氨基酸-tRNA与30S和50S亚基都发生交联，但fMet-tRNAfMet仅与50S亚基交联。在每种情况下，都发现tRNA的交联不仅发生在核糖体蛋白上，也发生在核糖体RNA（rRNA）上。共价复合物似乎足够稳定，能够鉴定出所涉及的蛋白质或rRNA序列。