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Structural analysis of the interaction of the tRNA modifying enzymes Tgt and QueA with a substrate tRNA
Author(s) -
Mueller Stefan O.,
Slany Robert K.
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)00169-a
Subject(s) - transfer rna , rnase p , biochemistry , biology , enzyme , endoribonuclease , escherichia coli , rna , gene
The enzymes tRNA guanine‐transglycosylase (Tgt) and S ‐adenosylmethionine : tRNA ribosyltransferase‐isomerase (QueA) participate in the biosynthesis of the hypermodified tRNA nucleoside queuosine (Q) in Escherichia coli . Here we show by HPLC analysis and gel retardation that both enzymes interact with an in vitro transcribed tRNA Asp from yeast, specifically modified with a Q precursor molecule. RNase I footprinting experiments showed strong protein tRNA contacts in the anticodon stem‐loop and a minor interaction with the dihydrouridine loop. This suggests that all identity elements for the recognition of Q‐specific tRNAs are clustered in the anticodon region and explains earlier results that both enzymes accept a RNA microhelix with the sequence of an anticodon stem‐loop as substrate.