Abstract
Previous sequence analysis of the Neurospora oli2 (ATP6) mitochondrial gene suggested that, in addition to a typical Group-I intron, it contained an unusual, mostly-palindromic, 93-nucleotide intron. We report here revisions of the nucleotide sequence and analysis of the size and sequence of reverse-transcriptase PCR products that show: (1) the Group-I intron splice sites are located as predicted by previous DNA sequence analysis; (2) the putative 93-nt intron is not excised from the mature mRNA, and most of this sequence is actually in the 5′ untranslated region. We conclude that the Neurospora ATP6 gene contains only one intro. Analysis of the cDNA sequence also confirms the non-universal nature of the Neurospora mitochondrial genetic code: a TGA codon inferred from the DNA sequence is present as UGA in the mRNA. This provides direct evidence that this codon is not altered, for example by RNA editing, to conform to the universal code.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bittner-Eddy, Monroy AF, Brambl R (1994) Expression of mitochondrial genes in the germinating conidia of Neurospora crassa. J Mol Biol (in press)
Burggraf S, Larsen N, Woese CR, Stetter KO (1993) An intron within the 16s ribosomal RNA gene of the archaeon Pyrobaculum aerophilum. Proc Natl Acad Sci USA 90:2547–2550
Cech TR (1988) Conserved sequences and structures of group I introns: building an active site for RNA catalysis — a review. Gene 73:259–271
Cech TR (1990) Splicing of Group-I introns. Annu Rev Biochem 59:543–568
Christopher DA, Hallick RB (1989) Euglena gracilis chloroplast ribosomal protein operon: a new chloroplast gene for ribosomal protein L5 and description of a novel organelle intron category designated group III. Nucleic Acids Res 17:7591–7608
Collins R (1990) Neurospora crassa laboratory strain 74-OR23-1A: mitochondrial genes. In: O'Brien SJ (ed) Genetic maps, 5th edn. Cold Spring Harbor Press, Cold Spring Harbor, New York, pp 3.19–3.21
Cummings DJ, Domenico JM (1988) Sequence analysis of mitochondrial DNA from Podospora anserina. Pervasiveness of a class-I intron in three separate genes. J Mol Biol 204:815–839
Davis RW, deSerres F (1970) Genetic and microbiological research techniques for Neurospora crassa. Methods Enzymol 17A: 70–143
Fox TD (1987) Natural variation in the genetic code. Annu Rev Genet 21:67–91
Garriga G, Bertrand H, Lambowitz AM (1984) RNA splicing in Neurospora mitochondria: nuclear mutants defective in both splicing and 3′-end synthesis of the large rRNA. Cell 36:623–634
Grisi E, Brown TA, Waring RB, Scazzocchio C, Davies RW (1982) Nucleotide sequence of a region of the mitochondrial genome of Aspergillus nidulans including the gene for ATPase 6. Nucleic Acids Res 10:3531–3539
Heckman JE, Sarnoff J, Alzner-DeWeerd B, Yin S, RajBhandary UL (1980) Novel features in the genetic code and codon reading patterns in Neurospora crassa mitochondria based on the sequences of six mitochondrial tRNAs. Proc Natl Acad Sci USA 77: 3159–3163
Lambowitz AM (1979) Preparation and analysis of mitochondrial ribosomes. Methods Enzymol 59:421–433
Lonergan KM, Gray MW (1993) Editing of transfer RNAs in Acanthamoeba castellanii mitochondria. Science 259:812–816
Macino G, Tzagoloff A (1980) Assembly of the mitochondrial membrane system: sequence analysis of a yeast mitochondrial ATPase gene containing oli-2 and oli-4 loci. Cell 20:507–517
Michel F, Umesono K, Ozeki H (1989) Comparative and functional anatomy of group II catalytic introns. Gene 82:5–30
Mizusawa S, Nishimura S, Seela F (1986) Improvement of the dideoxy chain-termination method of DNA sequencing by the use of deoxy-7-deazaguanosine triphosphate in place of dGTP. Nucleic Acids Res 14:1319–1324
Morelli G, Macino G (1984) Two intervening sequences in the ATPase subunit 6 gene of Neurospora crassa. A short intron (93 base-pairs) and a long intron that is stable after excision. J Mol Biol 178:491–507
Netzker R, Kochel HG, Basak N, Kuntzel H (1982) Nucleotide sequence of Aspergillus nidulans mitochondrial genes coding for ATPase subunit 6, cytochrome oxidase subunit 3, seven unidentified proteins, four tRNAs and L-rRNA. Nucleic Acids Res 10: 4783–4794
Newmeyer D, Perkins DD, Barry EG (1987) An annotated pedigree of Neurospora crassa laboratory wild-types, showing the probable origin of the nucleous satellite and showing that certain stocks are not authentic. Neurospora Newslett 25:46–51
Osawa S, Jukes TH, Watanabe K, Muto A (1992) Recent evidence for evolution of the genetic code. Microbiol Rev 56:229–264
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:77–90
Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Huguchi R, Horn GT, Mullis KB, Ehrlich HA (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
Taylor JW, Smolich BD (1985) Molecular cloning and physical mapping of the Neurospora crassa 74-OR23-1A mitochondrial genome Curr Genet 9:597–603
Wissinger B, Brennicke A, Schuster W (1992) Regenerating good sense: RNA editing and trans splicing in plant mitochondria. Trends in Genet 8:322–328
Author information
Authors and Affiliations
Additional information
Communicated by R.W. Lee
Rights and permissions
About this article
Cite this article
Collins, R.A., Olive, J.E. Revision of the nucleotide sequence and RNA splicing pathway of the Neurospora mitochondrial gene encoding ATPase subunit 6. Curr Genet 25, 514–518 (1994). https://doi.org/10.1007/BF00351671
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00351671