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Nucleic Acids Res. 1992 April 11; 20(7): 1747–1754.
PMCID: PMC312266

A maturase-encoding group IIA intron of yeast mitochondria self-splices in vitro.

Abstract

Intron 1 of the coxI gene of yeast mitochondrial DNA (aI1) is a group IIA intron that encodes a maturase function required for its splicing in vivo. It is shown here to self-splice in vitro under some reaction conditions reported earlier to yield efficient self-splicing of group IIB introns of yeast mtDNA that do not encode maturase functions. Unlike the group IIB introns, aI1 is inactive in 10 mM Mg2+ (including spermidine) and requires much higher levels of Mg2+ and added salts (1M NH4Cl or KCl or 2M (NH4)2SO4) for ready detection of splicing activity. In KCl-stimulated reactions, splicing occurs with little normal branch formation; a post-splicing reaction of linear excised intron RNA that forms shorter lariat RNAs with branches at cryptic sites was evident in those samples. At low levels of added NH4Cl or KCl, the precursor RNA carries out the first reaction step but appears blocked in the splicing step. AI1 RNA is most reactive at 37-42 degrees C, as compared with 45 degrees C for the group IIB introns; and it lacks the KCl- or NH4Cl-dependent spliced-exon reopening reaction that is evident for the self-splicing group IIB introns of yeast mitochondria. Like the group IIB intron aI5 gamma, the domain 4 of aI1 can be largely deleted in cis, without blocking splicing; also, trans-splicing of half molecules interrupted in domain 4 occurs. This is the first report of a maturase-encoding intron of either group I or group II that self-splices in vitro.

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Selected References

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  • Kück U, Godehardt I, Schmidt U. A self-splicing group II intron in the mitochondrial large subunit rRNA (LSUrRNA) gene of the eukaryotic alga Scenedesmus obliquus. Nucleic Acids Res. 1990 May 11;18(9):2691–2697. [PMC free article] [PubMed]
  • Schmidt U, Riederer B, Mörl M, Schmelzer C, Stahl U. Self-splicing of the mobile group II intron of the filamentous fungus Podospora anserina (COI I1) in vitro. EMBO J. 1990 Jul;9(7):2289–2298. [PubMed]
  • Garriga G, Lambowitz AM. RNA splicing in neurospora mitochondria: self-splicing of a mitochondrial intron in vitro. Cell. 1984 Dec;39(3 Pt 2):631–641. [PubMed]
  • van der Horst G, Tabak HF. Self-splicing of yeast mitochondrial ribosomal and messenger RNA precursors. Cell. 1985 Apr;40(4):759–766. [PubMed]
  • Gott JM, Shub DA, Belfort M. Multiple self-splicing introns in bacteriophage T4: evidence from autocatalytic GTP labeling of RNA in vitro. Cell. 1986 Oct 10;47(1):81–87. [PubMed]
  • Tabak HF, Van der Horst G, Kamps AM, Arnberg AC. Interlocked RNA circle formation by a self-splicing yeast mitochondrial group I intron. Cell. 1987 Jan 16;48(1):101–110. [PubMed]
  • Muscarella DE, Vogt VM. A mobile group I intron in the nuclear rDNA of Physarum polycephalum. Cell. 1989 Feb 10;56(3):443–454. [PubMed]
  • Garriga G, Lambowitz AM. RNA splicing in Neurospora mitochondria. The large rRNA intron contains a noncoded, 5'-terminal guanosine residue. J Biol Chem. 1983 Dec 25;258(24):14745–14748. [PubMed]
  • Garriga G, Lambowitz AM. Protein-dependent splicing of a group I intron in ribonucleoprotein particles and soluble fractions. Cell. 1986 Aug 29;46(5):669–680. [PubMed]
  • Gampel A, Tzagoloff A. In vitro splicing of the terminal intervening sequence of Saccharomyces cerevisiae cytochrome b pre-mRNA. Mol Cell Biol. 1987 Jul;7(7):2545–2551. [PMC free article] [PubMed]
  • Partono S, Lewin AS. Autocatalytic activities of intron 5 of the cob gene of yeast mitochondria. Mol Cell Biol. 1988 Jun;8(6):2562–2571. [PMC free article] [PubMed]
  • McGraw P, Tzagoloff A. Assembly of the mitochondrial membrane system. Characterization of a yeast nuclear gene involved in the processing of the cytochrome b pre-mRNA. J Biol Chem. 1983 Aug 10;258(15):9459–9468. [PubMed]
  • Gampel A, Nishikimi M, Tzagoloff A. CBP2 protein promotes in vitro excision of a yeast mitochondrial group I intron. Mol Cell Biol. 1989 Dec;9(12):5424–5433. [PMC free article] [PubMed]
  • Cech TR. Conserved sequences and structures of group I introns: building an active site for RNA catalysis--a review. Gene. 1988 Dec 20;73(2):259–271. [PubMed]
  • Michel F, Umesono K, Ozeki H. Comparative and functional anatomy of group II catalytic introns--a review. Gene. 1989 Oct 15;82(1):5–30. [PubMed]
  • Cech TR. Self-splicing of group I introns. Annu Rev Biochem. 1990;59:543–568. [PubMed]
  • Perlman PS. Genetic analysis of RNA splicing in yeast mitochondria. Methods Enzymol. 1990;181:539–558. [PubMed]
  • Lambowitz AM, Perlman PS. Involvement of aminoacyl-tRNA synthetases and other proteins in group I and group II intron splicing. Trends Biochem Sci. 1990 Nov;15(11):440–444. [PubMed]
  • Peebles CL, Perlman PS, Mecklenburg KL, Petrillo ML, Tabor JH, Jarrell KA, Cheng HL. A self-splicing RNA excises an intron lariat. Cell. 1986 Jan 31;44(2):213–223. [PubMed]
  • van der Veen R, Arnberg AC, van der Horst G, Bonen L, Tabak HF, Grivell LA. Excised group II introns in yeast mitochondria are lariats and can be formed by self-splicing in vitro. Cell. 1986 Jan 31;44(2):225–234. [PubMed]
  • Schmelzer C, Schweyen RJ. Self-splicing of group II introns in vitro: mapping of the branch point and mutational inhibition of lariat formation. Cell. 1986 Aug 15;46(4):557–565. [PubMed]
  • Carignani G, Groudinsky O, Frezza D, Schiavon E, Bergantino E, Slonimski PP. An mRNA maturase is encoded by the first intron of the mitochondrial gene for the subunit I of cytochrome oxidase in S. cerevisiae. Cell. 1983 Dec;35(3 Pt 2):733–742. [PubMed]
  • Carignani G, Netter P, Bergantino E, Robineau S. Expression of the mitochondrial split gene coding for cytochrome oxidase subunit I in S. cerevisiae: RNA splicing pathway. Curr Genet. 1986;11(1):55–63. [PubMed]
  • Hudspeth ME, Shumard DS, Tatti KM, Grossman LI. Rapid purification of yeast mitochondrial DNA in high yield. Biochim Biophys Acta. 1980 Dec 11;610(2):221–228. [PubMed]
  • Bonitz SG, Coruzzi G, Thalenfeld BE, Tzagoloff A, Macino G. Assembly of the mitochondrial membrane system. Structure and nucleotide sequence of the gene coding for subunit 1 of yeast cytochrme oxidase. J Biol Chem. 1980 Dec 25;255(24):11927–11941. [PubMed]
  • de Zamaroczy M, Bernardi G. The primary structure of the mitochondrial genome of Saccharomyces cerevisiae--a review. Gene. 1986;47(2-3):155–177. [PubMed]
  • Jarrell KA, Peebles CL, Dietrich RC, Romiti SL, Perlman PS. Group II intron self-splicing. Alternative reaction conditions yield novel products. J Biol Chem. 1988 Mar 5;263(7):3432–3439. [PubMed]
  • Sharp PA, Berk AJ, Berget SM. Transcription maps of adenovirus. Methods Enzymol. 1980;65(1):750–768. [PubMed]
  • Lane DJ, Pace B, Olsen GJ, Stahl DA, Sogin ML, Pace NR. Rapid determination of 16S ribosomal RNA sequences for phylogenetic analyses. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6955–6959. [PubMed]
  • England TE, Bruce AG, Uhlenbeck OC. Specific labeling of 3' termini of RNA with T4 RNA ligase. Methods Enzymol. 1980;65(1):65–74. [PubMed]
  • Müller MW, Schweyen RJ, Schmelzer C. Selection of cryptic 5' splice sites by group II intron RNAs in vitro. Nucleic Acids Res. 1988 Aug 11;16(15):7383–7395. [PMC free article] [PubMed]
  • Ruskin B, Green MR. An RNA processing activity that debranches RNA lariats. Science. 1985 Jul 12;229(4709):135–140. [PubMed]
  • Jarrell KA, Dietrich RC, Perlman PS. Group II intron domain 5 facilitates a trans-splicing reaction. Mol Cell Biol. 1988 Jun;8(6):2361–2366. [PMC free article] [PubMed]

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