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. 1984 Feb;3(2):377–382. doi: 10.1002/j.1460-2075.1984.tb01815.x

Nucleotide sequence of the cloned mRNA and gene of the ADP/ATP carrier from Neurospora crassa.

H Arends, W Sebald
PMCID: PMC557353  PMID: 6325169

Abstract

A cDNA complementary to the mRNA of the ADP/ATP carrier from Neurospora crassa was identified among ordered cDNA clones by hybridizing total polyadenylated RNA to pools of 96 cDNA recombinant plasmids and subsequent cell-free translation of hybridization-selected mRNA. Further carrier cDNAs were found by colony filter hybridization at a frequency of 0.2-0.3%. The gene of the carrier was cloned and isolated on a 4.6-kbp EcoRI fragment of total Neurospora DNA, and the start of the mRNA was determined by S1 nuclease mapping. From the nucleotide sequence of the cDNA and the genomic DNA, the primary structure of the gene, of the mRNA and of the ADP/ATP carrier protein could be deduced. The gene occurs in a single copy in the genome and related genes are absent. It contains two short introns, and a pyrimidine-rich promoter region. The mRNA has a 46-bp 5' end and a 219-bp 3' end. There is an open reading frame coding for the 313 amino acid residues of the Neurospora carrier protein. The amino acid sequence is homologous in 148 positions with the established primary structure of the beef heart carrier.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alton N. K., Buxton F., Patel V., Giles N. H., Vapnek D. 5'-Untranslated sequences of two structural genes in the qa gene cluster of Neurospora crassa. Proc Natl Acad Sci U S A. 1982 Mar;79(6):1955–1959. doi: 10.1073/pnas.79.6.1955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aquila H., Misra D., Eulitz M., Klingenberg M. Complete amino acid sequence of the ADP/ATP carrier from beef heart mitochondria. Hoppe Seylers Z Physiol Chem. 1982 Mar;363(3):345–349. [PubMed] [Google Scholar]
  3. Gross-Bellard M., Oudet P., Chambon P. Isolation of high-molecular-weight DNA from mammalian cells. Eur J Biochem. 1973 Jul 2;36(1):32–38. doi: 10.1111/j.1432-1033.1973.tb02881.x. [DOI] [PubMed] [Google Scholar]
  4. Hackenberg H., Riccio P., Klingenberg M. The biosynthesis of the mitochondrial ADP, ATP translocator. Eur J Biochem. 1978 Aug 1;88(2):373–378. doi: 10.1111/j.1432-1033.1978.tb12458.x. [DOI] [PubMed] [Google Scholar]
  5. Hallermayer G., Zimmermann R., Neupert W. Kinetic studies on the transport of cytoplasmically synthesized proteins into the mitochondria in intact cells of Neurospora crassa. Eur J Biochem. 1977 Dec;81(3):523–532. doi: 10.1111/j.1432-1033.1977.tb11978.x. [DOI] [PubMed] [Google Scholar]
  6. Jeffreys A. J., Flavell R. A. A physical map of the DNA regions flanking the rabbit beta-globin gene. Cell. 1977 Oct;12(2):429–439. doi: 10.1016/0092-8674(77)90119-2. [DOI] [PubMed] [Google Scholar]
  7. Klingenberg M. The ADP-ATP translocation in mitochondria, a membrane potential controlled transport. J Membr Biol. 1980 Sep 30;56(2):97–105. doi: 10.1007/BF01875961. [DOI] [PubMed] [Google Scholar]
  8. Langford C. J., Gallwitz D. Evidence for an intron-contained sequence required for the splicing of yeast RNA polymerase II transcripts. Cell. 1983 Jun;33(2):519–527. doi: 10.1016/0092-8674(83)90433-6. [DOI] [PubMed] [Google Scholar]
  9. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  10. O'Malley K., Pratt P., Robertson J., Lilly M., Douglas M. G. Selection of the nuclear gene for the mitochondrial adenine nucleotide translocator by genetic complementation of the op1 mutation in yeast. J Biol Chem. 1982 Feb 25;257(4):2097–2103. [PubMed] [Google Scholar]
  11. Perkins D. D., Radford A., Newmeyer D., Björkman M. Chromosomal loci of Neurospora crassa. Microbiol Rev. 1982 Dec;46(4):426–570. doi: 10.1128/mr.46.4.426-570.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Rüther U., Koenen M., Otto K., Müller-Hill B. pUR222, a vector for cloning and rapid chemical sequencing of DNA. Nucleic Acids Res. 1981 Aug 25;9(16):4087–4098. doi: 10.1093/nar/9.16.4087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Saraste M., Walker J. E. Internal sequence repeats and the path of polypeptide in mitochondrial ADP/ATP translocase. FEBS Lett. 1982 Aug 2;144(2):250–254. doi: 10.1016/0014-5793(82)80648-0. [DOI] [PubMed] [Google Scholar]
  14. Schechtman M. G., Yanofsky C. Structure of the trifunctional trp-1 gene from Neurospora crassa and its aberrant expression in Escherichia coli. J Mol Appl Genet. 1983;2(1):83–99. [PubMed] [Google Scholar]
  15. Sebald W., Graf T., Lukins H. B. The dicyclohexylcarbodiimide-binding protein of the mitochondrial ATPase complex from Neurospora crassa and Saccharomyces cerevisiae. Identification and isolation. Eur J Biochem. 1979 Feb 1;93(3):587–599. doi: 10.1111/j.1432-1033.1979.tb12859.x. [DOI] [PubMed] [Google Scholar]
  16. Viebrock A., Perz A., Sebald W. The imported preprotein of the proteolipid subunit of the mitochondrial ATP synthase from Neurospora crassa. Molecular cloning and sequencing of the mRNA. EMBO J. 1982;1(5):565–571. doi: 10.1002/j.1460-2075.1982.tb01209.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Vignais P. V. Molecular and physiological aspects of adenine nucleotide transport in mitochondria. Biochim Biophys Acta. 1976 Apr 30;456(1):1–38. doi: 10.1016/0304-4173(76)90007-0. [DOI] [PubMed] [Google Scholar]
  18. Weaver R. F., Weissmann C. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates. Nucleic Acids Res. 1979 Nov 10;7(5):1175–1193. doi: 10.1093/nar/7.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Woudt L. P., Pastink A., Kempers-Veenstra A. E., Jansen A. E., Mager W. H., Planta R. J. The genes coding for histone H3 and H4 in Neurospora crassa are unique and contain intervening sequences. Nucleic Acids Res. 1983 Aug 25;11(16):5347–5360. doi: 10.1093/nar/11.16.5347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Zimmerman R., Paluch U., Sprinzl M., Neupert W. Cell-free synthesis of the mitochondrial ADP/ATP carrier protein of Neurospora crassa. Eur J Biochem. 1979 Sep;99(2):247–252. doi: 10.1111/j.1432-1033.1979.tb13251.x. [DOI] [PubMed] [Google Scholar]
  21. Zwizinski C., Schleyer M., Neupert W. Transfer of proteins into mitochondria. Precursor to the ADP/ATP carrier binds to receptor sites on isolated mitochondria. J Biol Chem. 1983 Apr 10;258(7):4071–4074. [PubMed] [Google Scholar]
  22. von Heijne G. Membrane proteins: the amino acid composition of membrane-penetrating segments. Eur J Biochem. 1981 Nov;120(2):275–278. doi: 10.1111/j.1432-1033.1981.tb05700.x. [DOI] [PubMed] [Google Scholar]
  23. von Jagow G., Weiss H., Klingenberg M. Comparison of the respiratory chain of Neurospora crassa wild type and the mi-mutants mi-1 and mi-3. Eur J Biochem. 1973 Feb 15;33(1):140–157. doi: 10.1111/j.1432-1033.1973.tb02665.x. [DOI] [PubMed] [Google Scholar]

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