RNA-PROTEIN CROSS-LINKING IN ESCHERICHIA-COLI 30S RIBOSOMAL-SUBUNITS - PRECISE LOCALIZATION OF THE NUCLEOTIDE IN 16S RNA WHICH IS COUPLED TO PROTEIN-S7 BY ULTRAVIOLET-IRRADIATION

被引:75
作者
ZWIEB, C
BRIMACOMBE, R
机构
[1] Max-Planck-Institut fur Molekulare Genetik, Abteilung Wittmann, Berlin-Dahlem
关键词
D O I
10.1093/nar/6.5.1775
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
It is well established that when E. coil 30s ribosomal subunits are irradiated with ultraviolet light under mild conditions a specific cross-link is formed between protein S7 and the 16s RNA. Methodology is presented for the analysis of the single nucleotide residue concerned in this cross-link. Firstly, the identity of the ribonuclease T1 octanucleotide attached to S7 is confirmed by a new method, which involves isolation and analysis of S7-polynucleotide complexes containing 30 - 40 nucleotides. Secondly, the isolated S7-octanucleotide complex is digested successively with ribonuclease A, proteinase K and ribonuclease T2, and the nucleotides liberated are identified. The results show unambiguously that uridine residue number 1239 in the 16s RNA sequence is cross-linked to protein S7. © 1979 Information Retrieval Limited.
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页码:1775 / 1790
页数:16
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