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An engineered class I transfer RNA with a class II tertiary fold

Published online by Cambridge University Press:  01 March 1999

T. AMAR NISSAN ,
BRYANT OLIPHANT  and
JOHN J. PERONA
T. AMAR NISSAN
Affiliation:
Department of Chemistry, and Interdepartmental Program in Biochemistry and Molecular Biology, University of California at Santa Barbara, Santa Barbara, California 93106-9510, USA
BRYANT OLIPHANT
Affiliation:
Department of Chemistry, and Interdepartmental Program in Biochemistry and Molecular Biology, University of California at Santa Barbara, Santa Barbara, California 93106-9510, USA
JOHN J. PERONA
Affiliation:
Department of Chemistry, and Interdepartmental Program in Biochemistry and Molecular Biology, University of California at Santa Barbara, Santa Barbara, California 93106-9510, USA
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Abstract

Structure-based engineering of the tertiary fold of Escherichia coli tRNA2Gln has enabled conversion of this transfer RNA to a class II structure while retaining recognition properties of a class I glutamine tRNA. The new tRNA possesses the 20-nt variable stem-loop of Thermus thermophilus tRNASer. Enlargement of the D-loop appears essential to maintaining a stable tertiary structure in this species, while rearrangement of a base triple in the augmented D-stem is critical for efficient glutaminylation. These data provide new insight into structural determinants distinguishing the class I and class II tRNA folds, and demonstrate a marked sensitivity of glutaminyl-tRNA synthetase to alteration of tRNA tertiary structure.

Keywords

aminoacyl-tRNA synthetasesextra armRNA engineeringRNA structuretRNA identity
Type
Research Article
Information
RNA , Volume 5 , Issue 3 , March 1999 , pp. 434 - 445
Copyright
1999 RNA Society

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