Summary
In the filamentous fungus Podospora anserina, the amplification as circular DNA molecules of the first intron (intron α) of the CO1 mitochondrial gene, encoding the cytochrome oxidase subunit 1, is known to be strongly associated with aging of strains. In this study we have attempted to detect the protein potentially encoded by the open reading frame (ORF) contained in this intron. This was done by the Western blot technique using specific antisera raised against three polypeptides encoded by three non-overlapping fragments of this ORF adapted to the universal code and overexpressed in Escherichia coli. We examined about thirty independent subclones of Podospora derived from two different geographic races (A, s), using wild-type and mutant strains, young and senescent cultures. A 100 kDa polypeptide, encoded by the class II intron α, was detected in five senescent subclones which all showed strong amplification of the intronic α sequence (Sen DNA α).
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Belcour L, Vierny C (1986) Variable DNA splicing sites of a mitochondrial intron: relationship to the senescence process in Podospora. EMBO J 5:609–614
Belcour L, Begel O, Mossé MO, Vierny C (1981) Mitochondrial DNA amplification in senescent cultures of Podospora anserina: variability between the retained amplified sequences. Curr Genet 3: 13–21
Belcour L, Koll F, Vierny C, Sainsard-Chanet A, Begel O (1986) Are proteins encoded in mitochondrial introns involved in the process of senescence in the fungus Podospora anserina? In: Courtois Y, Faucheux B, Forette B, Knook DL, Tréton JA (eds) Modern trands in aging research, John Libbey, Eurotext London Paris, vol 147, pp 63–71
Carginani G, Groudinski O, Frezza D, Schiavon E, Bergantino E, Slonimski PP (1983) An mRNA maturase is encoded by the first intron of the mitochondrial gene for the subunit 1 of cytochrome oxidase in S. cerevisiae. Cell 35:733–742
Carter P, Bedouelle H, Winter G (1986) Improved oligonucleotide site-directed mutagenesis using M13 vectors. Nucleic Acids Res 13:4431–4443
Colleaux L, d'Auriol L, Betermier M, Cottarel G, Jacquier A, Galibert F, Dujon B (1986) Universal code equivalent of a yeast mitochondrial intron reading frame is expressed in E. coli as a specific double strand endonuclease. Cell 44:521–533
Cummings DJ, Wright RM (1983) DNA sequence of the excision sites of a mitochondrial plasmid from senescent Podospora anserina. Nucleic Acids Res 11:7–15
Cummings DJ, Belcour L, Grandchamp C (1979) Mitochondrial DNA from Podospora anserina. Isolation and characterization. Mol Gen Genet 171:229–238
Cummings DJ, MacNeil IA, Domenico J, Matsuura ET (1985) Excision-amplification of mitochondrial DNA during senescence in Podospora anserina; DNA sequence analysis of three unique “plasmids”. J Mol Biol 185:659–680
Delahodde A, Banroques J, Bécam AM, Goguel V, Perea J, Schroeder R, Jacq C (1985) Purification of yeast bl14 mRNA maturase from E. coli and from yeast. Nucleic acids binding properties. In: Quagliariello E, Slater EC, Palmieri F, Saccone C, Kroon AM (eds) Achievements and perspectives of mitochondria research, vol 2. Elsevier Science Publishers, Amsterdam, pp 79–88
Delahodde A, Goguel V, Becam AM, Creusot F, Perea J, Bantoques J, Jacq C (1989) Site-specific DNA endonuclease and RNA maturase activities of two homologous intron-encoded proteins from yeast mitochondria. Cell 56:431–441
Dequart-Chablat M, Coppin-Raynal E, Picard-Bennoun M, Madjar JJ (1986) At least seven ribosomal proteins are involved in the control of translational accuracy in a eukaryotic organism. J Mol Biol 190:167–175
Dujon B, Colleaux L, Jacquier A, Michel F, Monteihet C (1986) Mitochondrial introns as mobile genetic elements: the role of intron-encoded proteins. In: Wickner RB, Hinnebusch A, Lambowitz AM, Gunsalus IC, Hollaender A (eds) Extrachromosomal elements in lower eukaryotes. Plenum, New York, pp 5–27
Goguel V, Bailone A, Devoret R, Jacq C (1989) The bl4 RNA mitochondrial maturase of Saccharomyces cerevisiae can stimulate intra-chromosomal recombination in Escherichia coli. Mol Gen Genet 216:70–74
Grosjean H, Fiers W (1982) Preferential codon usage in prokaryotic genes: the optimal codon-anticodon interaction energy and the selective codon usage in efficiently expressed genes. Gene 18:199–209
Herbert CJ, Labouesse M, Dujardin G, Slonimski PP (1988) The NAM2 proteins from S. cerevisiae and S. douglasii are mitochondrial leucyl-tRNA synthetases, and are involved in mRNA splicing. EMBO J 7:473–483
Jacq C, Lazowska J, Slonimski PP (1980) Sur un nouveau mécanisme de la régulation de l'expression génétique. CR Acad Sci Paris 290:89–92
Johnson DA, Gautsch JW, Sportsman JR, Elder JH (1984) Improved technique utilizing nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Anal Tech 1:3–8
Kück U (1988) Putative nucleic acid binding ‘fingers’ at the carboxyl terminus of polypeptides encoded from introns of organellar genes. Nucleic Acids Res 16:4729
Kück U (1989) Mitochondrial DNA rearrangements in Podospora anserina. Exp Mycol 13:111–120
Kück U, Stahl U, Esser K (1981) Plasmid-like DNA is part of mitochondrial DNA in Podospora anserina. Curr Genet 3:151–156
Lazowska J, Jacq C, Slonimski PP (1980) Sequence of introns and flanking exons in wild type and box 3 mutants reveals an interlaced splicing protein coded by an intron. Cell 22:333–348
Levra-Juillet E, Boulet A, Seraphin B, Simon M, Faye G (1989) Mitochondrial intron al1 and/or al2 are needed for the in vivo deletion of intervening sequences. Mol Gen Genet 217:168–171
Marcou D (1961) Notion de longévité et nature cytoplasmique du déterminant de la sénescence chez quelques champignons. Ann Sci Nat Bota 11:653–764
Matsuura ET, Domenico J, Cummings DJ (1986) An additional class II intron with homology to reverse transcriptase in rapidly senescing Podospora anserina. Curr Genet 10: 915–922
Meunier B, Tian GL, Macadre C, Slonimski PP, Lazowska J (1990) Group 11 introns transpose in yeast mitochondria. In: Quagliariello E, Papa S, Palmieri F, Saccone C (eds) Structure, function and biogenesis of energy transfer systems. Elsevier Science Publishers, Amsterdam, pp 169–173
Michel F, Dujon B (1983) Conservation of RNA secondary structures in two intron families including mitochondrial- chloroplast- and nuclear-encoded members. EMBO J 2:33–38
Michel F, Lang BF (1985) Mitochondrial class II introns encode proteins related to the reverse transcriptase of retroviruses. Nature 316:641–643
Osiewacz HD, Esser K (1984) The mitochondrial plasmid of Podospora anserina: A mobile intron of a mitochondrial gene. Curr Genet 8:229–305
Peebles CL, Perlman PS, Mecklenburg KL, Petrillo ML, Tabor JH, Jarell KA, Cheng HL (1986) A self-splicing RNA excises an intron lariat. Cell 44:213–223
Sainsard-Chanet A, Begel O (1990) Insertion of an LrDNA gene fragment and of filler DNA at a mitochondrial exon-intron junction in Podospora. Nucleic Acids Res 18:779–783
Schmelzer C, Schweyen RJ (1986) Self-splicing of group II introns in vitro: mapping of the branch point and mutational inhibition of lariat formation. Cell 46:557–565
Schmidt U, Kosack M, Stahl U (1987) Lariat RNA of a group II intron in a filamentous fungus. Curr Genet 12:291–295
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Towbin H, Staehlin T, Gordon J (1976) Electrophoretic transfer of protein from acrylamide gels to nitocellulose sheets: procedure and application. Proc Natl Acad Sci USA 76:4350–4354
van der Veen R, Arnberg AC, van der Horst G, Bonen L, Tabak HF, Grivell LA (1986) Excised group II introns in yeast mitochondria are lariats and can be formed by self-splicing in vitro. Cell 44:225–234
Vierny C, Keller AM, Begel O, Belcour L (1982) A sequence of mitochondrial DNA is associated with the onset of senescence in a fungus. Nature 297:157–159
Wenzlau JM, Saldanha RJ, Butow RA, Perlman PS (1989) A latent intron-encoded maturase is also an endonuclease needed for intron mobility. Cell 56:421–430
Wright RM, Horum MA, Cummings DJ (1982) Are mitochondrial structural genes selectively amplified during senescence in Podospora anserina? Cell 29:505–515
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Communicated by W. Gajewski
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Sellem, C.H., Sainsard-Chanet, A. & Belcour, L. Detection of a protein encoded by a class II mitochondrial intron of Podospora anserina . Molec. Gen. Genet. 224, 232–240 (1990). https://doi.org/10.1007/BF00271556
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DOI: https://doi.org/10.1007/BF00271556