Article ID Journal Published Year Pages File Type
1994385 Methods 2008 10 Pages PDF
Abstract

Here we describe the many applications of acid urea polyacrylamide gel electrophoresis (acid urea PAGE) followed by Northern blot analysis to studies of tRNAs and aminoacyl-tRNA synthetases. Acid urea PAGE allows the electrophoretic separation of different forms of a tRNA, discriminated by changes in bulk, charge, and/or conformation that are brought about by aminoacylation, formylation, or modification of a tRNA. Among the examples described are (i) analysis of the effect of mutations in the Escherichia coli initiator tRNA on its aminoacylation and formylation; (ii) evidence of orthogonality of suppressor tRNAs in mammalian cells and yeast; (iii) analysis of aminoacylation specificity of an archaeal prolyl-tRNA synthetase that can aminoacylate archaeal tRNAPro with cysteine, but does not aminoacylate archaeal tRNACys with cysteine; (iv) identification and characterization of the AUA-decoding minor tRNAIle in archaea; and (v) evidence that the archaeal minor tRNAIle contains a modified base in the wobble position different from lysidine found in the corresponding eubacterial tRNA.

Related Topics
Life Sciences Biochemistry, Genetics and Molecular Biology Biochemistry
Authors
, ,