HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Guo, Q. B. Articles by Lambowitz, A. M. Search for Related Content PubMed PubMed Citation Articles by Guo, Q. B. Articles by Lambowitz, A. M. Social Bookmarking                      What's this?

J. Biol. Chem., Vol. 266, Issue 3, 1809-1819, Jan, 1991

Structural analysis of the Neurospora mitochondrial large rRNA intron and construction of a mini-intron that shows protein-dependent splicing

QB Guo, RA Akins, G Garriga and AM Lambowitz
Department of Molecular Genetics, Ohio State University, Columbus 43210.

The gene encoding the Neurospora mitochondrial large rRNA contains a single group I intron of 2.3 kilobases that is not self-splicing in vitro. We showed previously that the splicing of this intron in vivo and in vitro is dependent on the Neurospora cyt-18 protein, mitochondrial tyrosyl-tRNA synthetase. In the present work, we carried out further structural analysis of the intron and constructed mutant derivatives of it in order to identify features that are either required for splicing or prevent it from self-splicing. Previous studies showed that the intron contains a large hairpin structure near the 5' splice site. By mapping RNase III cleavage sites, we identified this hairpin structure as an extended P2 stem. We construct a mini- intron of 388 nucleotides by deleting the 426-amino acid intron open reading frame, most of the 5' intron hairpin, and all of L8. This mini- intron shows the same protein-dependent splicing as the full length intron, but is still not self-splicing. Further deletions, which remove all of P2 or all or part of P4, P6, P7, or P9, inactivate splicing, suggesting that an intact group I intron core structure is required. Strengthening the P1, P10, or P9.0 pairings did not enable the mini- intron to self-splice. Our findings indicate that the inability of the mitochondrial large rRNA intron to self-splice reflects deficiency of a structure or activity required for cleavage at the 5' splice site, either in the intron core itself or in the interaction between the core and the P1 stem.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				Q. Vicens, P. J. Paukstelis, E. Westhof, A. M. Lambowitz, and T. R. Cech Toward predicting self-splicing and protein-facilitated splicing of group I introns RNA, October 1, 2008; 14(10): 2013 - 2029. [Abstract] [Full Text] [PDF]

				P. Tijerina, H. Bhaskaran, and R. Russell Nonspecific binding to structured RNA and preferential unwinding of an exposed helix by the CYT-19 protein, a DEAD-box RNA chaperone PNAS, November 7, 2006; 103(45): 16698 - 16703. [Abstract] [Full Text] [PDF]

				C. A. Myers, B. Kuhla, S. Cusack, and A. M. Lambowitz tRNA-like recognition of group I introns by a tyrosyl-tRNA synthetase PNAS, February 14, 2002; (2002) 52596299. [Abstract] [Full Text] [PDF]

				B. Chen, A. R. Kubelik, S. Mohr, and C. A. Breitenberger Cloning and Characterization of the Neurospora crassa cyt-5 Gene J. Biol. Chem., March 15, 1996; 271(11): 6537 - 6544. [Abstract] [Full Text] [PDF]

				L. C. Shaw and A. S. Lewin Protein-induced Folding of a Group I Intron in Cytochrome b Pre-mRNA J. Biol. Chem., September 15, 1995; 270(37): 21552 - 21562. [Abstract] [Full Text] [PDF]

				C. Creutz The annexins and exocytosis Science, November 6, 1992; 258(5084): 924 - 931. [Abstract] [PDF]

				Q Guo and A M Lambowitz A tyrosyl-tRNA synthetase binds specifically to the group I intron catalytic core. Genes & Dev., August 1, 1992; 6(8): 1357 - 1372. [Abstract] [PDF]

				A Gampel and T R Cech Binding of the CBP2 protein to a yeast mitochondrial group I intron requires the catalytic core of the RNA. Genes & Dev., October 1, 1991; 5(10): 1870 - 1880. [Abstract] [PDF]

				J D Kittle, G Mohr, J A Gianelos, H Wang, and A M Lambowitz The Neurospora mitochondrial tyrosyl-tRNA synthetase is sufficient for group I intron splicing in vitro and uses the carboxy-terminal tRNA-binding domain along with other regions. Genes & Dev., June 1, 1991; 5(6): 1009 - 1021. [Abstract] [PDF]

				C. A. Myers, B. Kuhla, S. Cusack, and A. M. Lambowitz tRNA-like recognition of group I introns by a tyrosyl-tRNA synthetase PNAS, March 5, 2002; 99(5): 2630 - 2635. [Abstract] [Full Text] [PDF]