通过紫外线交联揭示mRNA在30S起始复合物中的核糖体定位。
Ribosomal localization of the mRNA in the 30S initiation complex as revealed by UV crosslinking.
作者信息
Brandt R, Gualerzi C O
机构信息
Max-Planck-Institut für Molekulare Genetik, Berlin, Germany.
出版信息
FEBS Lett. 1992 Oct 26;311(3):199-202. doi: 10.1016/0014-5793(92)81101-q.
Translation initiation complexes consisting of 30S ribosomal subunits, 32P-labelled mRNA (002 mRNA), fMet-tRNA and the three initiation factors were subjected to UV-crosslinking to determine the protein and rRNA neighbors of the bound mRNA by immunochemical methods and by nucleic acid hybridization techniques. The mRNA was found to be crosslinked to a specific region of the 16S rRNA spanning from nucleotide 418 to 615 and to ribosomal proteins S1 and S21 (the main targets), S3, S10, S12 and S14; a low level of crosslinking was also detected with S2, S7, S13, S18 and S19.
由30S核糖体亚基、32P标记的mRNA（002 mRNA）、甲硫氨酰-tRNA以及三种起始因子组成的翻译起始复合物进行紫外线交联，通过免疫化学方法和核酸杂交技术确定结合的mRNA的蛋白质和rRNA邻位。发现mRNA与16S rRNA从核苷酸418到615的特定区域以及核糖体蛋白S1和S21（主要靶点）、S3、S10、S12和S14交联；还检测到与S2、S7、S13、S18和S19的低水平交联。
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