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Research Article

Group II introns deleted for multiple substructures retain self-splicing activity.

J L Koch, S C Boulanger, S D Dib-Hajj, S K Hebbar, P S Perlman
J L Koch
Department of Molecular Genetics, Ohio State University, Columbus 43210.
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S C Boulanger
Department of Molecular Genetics, Ohio State University, Columbus 43210.
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S D Dib-Hajj
Department of Molecular Genetics, Ohio State University, Columbus 43210.
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S K Hebbar
Department of Molecular Genetics, Ohio State University, Columbus 43210.
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P S Perlman
Department of Molecular Genetics, Ohio State University, Columbus 43210.
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DOI: 10.1128/MCB.12.5.1950
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ABSTRACT

Group II introns can be folded into highly conserved secondary structures with six major substructures or domains. Domains 1 and 5 are known to play key roles in self-splicing, while the roles of domains 2, 3, 4, and 6 are less clear. A trans assay for domain 5 function has been developed which indicates that domain 5 has a binding site on the precursor RNA that is not predicted from any secondary structure element. In this study, the self-splicing group II intron 5 gamma of the coxI gene of yeast mitochondrial DNA was deleted for various intron domains, singly and in combinations. Those mutant introns were characterized for self-splicing reactions in vitro as a means of locating the domain 5 binding site. A single deletion of domain 2, 3, 4, or 6 does not block in vitro reactions at either splice junction, though the deletion of domain 6 reduces the fidelity of 3' splice site selection somewhat. Even the triple deletion lacking domains 2, 4, and 6 retains some self-splicing activity. The deletion of domains 2, 3, 4, and 6 blocks the reaction at the 3' splice junction but not at the 5' junction. From these results, we conclude that the binding site for domain 5 is within domain 1 and that the complex of 5' exon, domain 1, and domain 5 (plus short connecting sequences) constitutes the essential catalytic core of this intron.

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Group II introns deleted for multiple substructures retain self-splicing activity.
J L Koch, S C Boulanger, S D Dib-Hajj, S K Hebbar, P S Perlman
Molecular and Cellular Biology May 1992, 12 (5) 1950-1958; DOI: 10.1128/MCB.12.5.1950

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Group II introns deleted for multiple substructures retain self-splicing activity.
J L Koch, S C Boulanger, S D Dib-Hajj, S K Hebbar, P S Perlman
Molecular and Cellular Biology May 1992, 12 (5) 1950-1958; DOI: 10.1128/MCB.12.5.1950
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