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A top-half tDNA minihelix is a good substrate for the eubacterial CCA-adding enzyme

Published online by Cambridge University Press:  01 March 1998

PEI-YONG SHI
Affiliation:
Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06520-8024, USA
ALAN M. WEINER
Affiliation:
Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06520-8024, USA Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06520-8024, USA
NANCY MAIZELS
Affiliation:
Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06520-8024, USA Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06520-8024, USA
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Abstract

The CCA-adding enzyme [ATP(CTP):tRNA nucleotidyltransferase] catalyzes the addition and regeneration of the 3′-terminal CCA sequence of tRNAs. We show that the CCA-adding enzyme will specifically add a CCA terminus to synthetic full-length tDNA and to DNA oligonucleotides corresponding to the “top half” of tRNA — the acceptor stem and TψC stem-loop of tRNA. CCA addition to the top half tDNA minihelices requires a 2′ as well as a 3′ OH at the 3′ terminus of the tDNA. Addition also depends on the length of the base paired stem, and is facilitated by, but is not dependent upon, the presence of a TψC loop. These results provide further evidence for independent functions of the top and bottom halves of tRNA, and support the hypothesis that these two structurally distinct and functionally independent domains evolved independently.

Type
Research Article
Information
RNA , Volume 4 , Issue 3 , March 1998 , pp. 276 - 284
Copyright
© 1998 RNA Society

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