This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Koch, J L Articles by Perlman, P S Search for Related Content PubMed PubMed Citation Articles by Koch, J L Articles by Perlman, P S

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

Department of Molecular Genetics, Ohio State University, Columbus 43210.

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.

This article has been cited by other articles:

• Fedorova, O., Pyle, A. M. (2008). A conserved element that stabilizes the group II intron active site. RNA 14: 1048-1056 [Abstract] [Full Text]  
• de Lencastre, A., Pyle, A. M. (2008). Three essential and conserved regions of the group II intron are proximal to the 5'-splice site. RNA 14: 11-24 [Abstract] [Full Text]  
• SEETHARAMAN, M., ELDHO, N. V., PADGETT, R. A., DAYIE, K. T. (2006). Structure of a self-splicing group II intron catalytic effector domain 5: Parallels with spliceosomal U6 RNA. RNA 12: 235-247 [Abstract] [Full Text]  
• Su, L. J., Waldsich, C., Pyle, A. M. (2005). An obligate intermediate along the slow folding pathway of a group II intron ribozyme. Nucleic Acids Res 33: 6674-6687 [Abstract] [Full Text]  
• Huang, H.-R., Chao, M. Y., Armstrong, B., Wang, Y., Lambowitz, A. M., Perlman, P. S. (2003). The DIVa Maturase Binding Site in the Yeast Group II Intron aI2 Is Essential for Intron Homing but Not for In Vivo Splicing. Mol. Cell. Biol. 23: 8809-8819 [Abstract] [Full Text]  
• Field, D J, Friesen, J D (1996). Functionally redundant interactions between U2 and U6 spliceosomal snRNAs.. Genes Dev. 10: 489-501 [Abstract]  
• Chanfreau, G, Jacquier, A (1994). Catalytic site components common to both splicing steps of a group II intron. Science 266: 1383-1387 [Abstract]  
• Hong, L, Hallick, R B (1994). A group III intron is formed from domains of two individual group II introns.. Genes Dev. 8: 1589-1599 [Abstract]  
• Wise, J. (1993). Guides to the heart of the spliceosome. Science 262: 1978-1979  
• Wassarman, D., Steitz, J. (1992). Interactions of small nuclear RNA's with precursor messenger RNA during in vitro splicing. Science 257: 1918-1925 [Abstract]