Antibodies Against a Fused 'lacZ-yeast Mitochondrial Intron' Gene Product Allow Identification of the MRNA Maturase Encoded by the Fourth Intron of the Yeast Cob-box Gene

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1984 Jul 1

PMID 6430696

Citations 20

Authors

C Jacq

J Banroques

A M Becam

P P SLONIMSKI

N Guiso

A Danchin

Affiliations

Soon will be listed here.

Abstract

Several missense or nonsense mutations have been localized in the fourth intron open reading frame (ORF) of the yeast mitochondrial cytochrome b gene. These results and the phenotypes of mutants strongly suggested that a mRNA maturase, controlling the expression of both cytochrome b and cytochrome oxidase subunit I (COXI) genes, is encoded in this ORF. To investigate more directly the biosynthesis of mRNA maturase we raised antibodies against a part of the putative ORF translation product. For that purpose we inserted a fragment of the ORF sequence, in phase, into the C-terminal EcoRI site of lacZ gene. The hybrid gene was then expressed in Escherichia coli under the control of either the wild-type lac promoter or the thermoregulated lambda system PR/cI857. The hybrid protein was partially purified and antibodies were raised against it. These antibodies recognized a mitochondrially coded protein, p27, in intron mutants, whereas no such protein was detected in the wild-type cell. These results demonstrate that the p27 protein, previously shown to be associated with the mRNA maturase activity, is actually translated from the intron ORF. The autoregulated mRNA maturase synthesis model is discussed in relation to these results.

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References

Bonitz S, Berlani R, Coruzzi G, Li M, Macino G, Nobrega F . Codon recognition rules in yeast mitochondria. Proc Natl Acad Sci U S A. 1980; 77(6):3167-70. PMC: 349575. DOI: 10.1073/pnas.77.6.3167. View

Jacq C, Lazowska J, SLONIMSKI P . [New mechanism for regulation of genetic expression]. C R Seances Acad Sci D. 1980; 290(2):89-92. View

Bonitz S, Coruzzi G, Thalenfeld B, 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; 255(24):11927-41. View

Lazowska J, Jacq C, SLONIMSKI P . Sequence of introns and flanking exons in wild-type and box3 mutants of cytochrome b reveals an interlaced splicing protein coded by an intron. Cell. 1980; 22(2 Pt 2):333-48. DOI: 10.1016/0092-8674(80)90344-x. View

Dhawale S, Hanson D, Alexander N, Perlman P, Mahler H . Regulatory interactions between mitochondrial genes: interactions between two mosaic genes. Proc Natl Acad Sci U S A. 1981; 78(3):1778-82. PMC: 319217. DOI: 10.1073/pnas.78.3.1778. View

Burnette W . "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981; 112(2):195-203. DOI: 10.1016/0003-2697(81)90281-5. View

de la Salle H, Jacq C, SLONIMSKI P . Critical sequences within mitochondrial introns: pleiotropic mRNA maturase and cis-dominant signals of the box intron controlling reductase and oxidase. Cell. 1982; 28(4):721-32. DOI: 10.1016/0092-8674(82)90051-4. View

Netter P, Jacq C, Carignani G, SLONIMSKI P . Critical sequences within mitochondrial introns: cis-dominant mutations of the "cytochrome-b-like" intron of the oxidase gene. Cell. 1982; 28(4):733-8. DOI: 10.1016/0092-8674(82)90052-6. View

Dujardin G, Jacq C, SLONIMSKI P . Single base substitution in an intron of oxidase gene compensates splicing defects of the cytochrome b gene. Nature. 1982; 298(5875):628-32. DOI: 10.1038/298628a0. View

10.

Weiss-Brummer B, Rodel G, Schweyen R, Kaudewitz F . Expression of the split gene cob in yeast: evidence for a precursor of a "maturase" protein translated from intron 4 and preceding exons. Cell. 1982; 29(2):527-36. DOI: 10.1016/0092-8674(82)90169-6. View

11.

Hudspeth M, Ainley W, Shumard D, Butow R, Grossman L . Location and structure of the var1 gene on yeast mitochondrial DNA: nucleotide sequence of the 40.0 allele. Cell. 1982; 30(2):617-26. DOI: 10.1016/0092-8674(82)90258-6. View

12.

Anziano P, Hanson D, Mahler H, Perlman P . Functional domains in introns: trans-acting and cis-acting regions of intron 4 of the cob gene. Cell. 1982; 30(3):925-32. DOI: 10.1016/0092-8674(82)90297-5. View

13.

Waring R, Davies R, Scazzocchio C, Brown T . Internal structure of a mitochondrial intron of Aspergillus nidulans. Proc Natl Acad Sci U S A. 1982; 79(20):6332-6. PMC: 347115. DOI: 10.1073/pnas.79.20.6332. View

14.

Davies R, Waring R, Ray J, Brown T, Scazzocchio C . Making ends meet: a model for RNA splicing in fungal mitochondria. Nature. 1982; 300(5894):719-24. DOI: 10.1038/300719a0. View

15.

Michel F, Jacquier A, Dujon B . Comparison of fungal mitochondrial introns reveals extensive homologies in RNA secondary structure. Biochimie. 1982; 64(10):867-81. DOI: 10.1016/s0300-9084(82)80349-0. View

16.

Burke J, RajBhandary U . Intron within the large rRNA gene of N. crassa mitochondria: a long open reading frame and a consensus sequence possibly important in splicing. Cell. 1982; 31(3 Pt 2):509-20. DOI: 10.1016/0092-8674(82)90307-5. View

17.

Grossman A, Burgess R, Walter W, Gross C . Mutations in the Ion gene of E. coli K12 phenotypically suppress a mutation in the sigma subunit of RNA polymerase. Cell. 1983; 32(1):151-9. DOI: 10.1016/0092-8674(83)90505-6. View

18.

Hensgens L, Bonen L, de Haan M, Van Der Horst G, Grivell L . Two intron sequences in yeast mitochondrial COX1 gene: homology among URF-containing introns and strain-dependent variation in flanking exons. Cell. 1983; 32(2):379-89. DOI: 10.1016/0092-8674(83)90457-9. View

19.

Cech T, Tanner N, Tinoco Jr I, Weir B, Zuker M, Perlman P . Secondary structure of the Tetrahymena ribosomal RNA intervening sequence: structural homology with fungal mitochondrial intervening sequences. Proc Natl Acad Sci U S A. 1983; 80(13):3903-7. PMC: 394167. DOI: 10.1073/pnas.80.13.3903. View

20.

Leplatois P, Danchin A . Vectors for high conditional expression of cloned genes. Biochimie. 1983; 65(6):317-24. DOI: 10.1016/s0300-9084(83)80153-9. View