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Complete 5′ and 3′ end maturation of group II intron-containing tRNA precursors

Published online by Cambridge University Press:  07 February 2001

JÖRG VOGEL
Affiliation:
University of Uppsala, Institute of Cellular and Molecular Biology, Department of Microbiology, Husargatan 3, S-751 24 Uppsala, Sweden
WOLFGANG R. HESS
Affiliation:
Humboldt-University, Department of Biology, D-10115 Berlin, Germany
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Abstract

Higher plant chloroplasts provide the only experimentally validated example of functional tRNA genes that are disrupted by group II introns. Here, precursor transcripts for tRNAGly(UCC), tRNAVal(UAC), and tRNAAla(UGC) were investigated for processing of 5′ leader and 3′ trailer sequences in vivo. Use of intron-specific primer pairs and inclusion of a barley chloroplast splicing mutant specifically allowed us to evaluate the potential effect of intervening sequences that disrupt tRNA secondary and tertiary structures. The data suggest that (1) neither integrity of the dihydrouridine nor the anticodon domain is required for the nucleotidyltransferase-mediated addition of 3′-terminal CCA; (2) interruption of these two structural elements by group II introns does not interfere with nucleotide-specific 5′ maturation by RNase P; (3) processing intermediates of chloroplast tRNAs can be 3′ polyadenylated; and (4) plastid DNA-encoded proteins are not required for 3′ and 5′ maturation of plastid tRNAs.

Type
Research Article
Copyright
2001 RNA Society

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