Biopolym. Cell. 2006; 22(3):201-209.
http://dx.doi.org/10.7124/bc.000731
Structure and Function of Biopolymers
The isolation of histidine tRNA from Thermus thermophilus and the study of its primary structure and interaction sites with homologous aminoacyl-tRNA synthetase
1Gudzera O. I., 1Krikliviy I. A., 1Yaremchuk A. D., 1Tukalo M. A.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680

Abstract

Histidine tRNAs (tRNAHis) are unique in possing an extra 5?-base (G-1) not found in other tRNAs. To study the molecular mechanisms of tRNAHis interaction with histidyl-tRNA synthetase, the method for purification of tRNAHis from Thermus thermophilus has been developed and tRNA1His has been sequenced. tRNAHis from Thermus thermophilus was isolated by combination of low-pressure benzoyl-DEAE- cellulose and DEAE Toyopearl 650 chromatographies with HPLC on DEAE 5PW and reversed phase columns. The nucleotide sequence of T. thermophilus tRNA1His has been determined by rapid gel-sequencing method. tRNA1His from T. thermophilus is different from those of E.coli at 23 positions. The sites of interaction of tRNAHis with histidyl-tRNA synthetase have been studied by the method of chemical modification with ethylnitrosourea. Histidyl-tRNA synthetase protects from modification following phospates: 8 – between acceptor and D-stems, 27, 28, 29 from 5?-end of anticodon stem, 34 – at the anticodon and phosphates 67, 68 from 3?-end of acceptor stem. All the protected sites of tRNAHis are found on one side of three-dimensional structure of tRNA where the variable stem is also located. D-stem is located on the opposite side and does not interact with the enzyme.
Keywords: tRNA, aminoacyl-tRNA synthetase, RNA-protein recognition, chemical modification
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