Abstract
Revertants were isolated from v-fos-transformed rat-1 cells cotransfected with a human cDNA expression library and a selectable marker (pMEX-neo). Molecular analysis of one clone, R2.2, suggested that the revertant phenotype resulted from the disruption of a transformation effector gene by the integration of the pMEX-neo plasmid. Genomic sequences flanking the plasmid integration site were cloned and used as probes in Northern blot analyses. A probe derived from sequences 5' to the integration site hybridized to a unique 1.2-kilobase mRNA and was used to isolate a 0.9-kilobase cDNA clone (fte-1). The open reading frame of the fte-1 cDNA predicts a highly basic protein that shows a remarkable level of similarity with two genes from Saccharomyces cerevisiae. One of these yeast genes contains an unidentified open reading frame and the other, MFT1, is a gene isolated from a yeast mutant that fails to import a fusion protein into mitochondria [Garrett, J. M., Singh, K. K., Vonder Haar, R. A. & Emr, S. D. (1991) Mol. Gen. Genet. 225, 483-491]. Expression of the fte-1 gene was induced approximately 5-fold in v-fos-transformed fibroblasts, but expression was reduced in clone R2.2 and in several independent revertant clones. Transfection of R2.2 cells with fte-1 expression vectors resulted in the reacquisition of a transformed phenotype. These results demonstrate that the mammalian homologue of a gene implicated in protein import into yeast mitochondria is a v-fos transformation effector gene.
MeSH Terms
Amino Acid Sequence
Animals
Base Sequence
Blotting, Southern
Carrier Proteins/genetics
Cell Line
Cell Line, Transformed
Cell Transformation, Viral/genetics
Cells, Cultured
Clone Cells
DNA/genetics,isolation & purification
Gene Library
Genes, Fungal
Genes, fos
Humans
Mitochondria/metabolism
Molecular Sequence Data
Open Reading Frames
Plasmids
RNA, Messenger/genetics
Rats
Restriction Mapping
Ribosomal Proteins
Saccharomyces cerevisiae/genetics,metabolism
Sequence Homology, Nucleic Acid
Transfection
Chemicals
Carrier Proteins
RNA, Messenger
RPS3A protein, human
Ribosomal Proteins
Rps3a protein, rat
DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kho C J
Division of Toxicology, Whitaker College of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge 02139.
Zarbl H
References (31)
31 references, click to expand
-
Biochemical transfer of single-copy eucaryotic genes using total cellular DNA as donor.
Cell. 1978 Jul;14(3):725-31
PMID: 210957
-
Isolation of Genes that Are Preferentially Expressed at the G(1)/S Boundary during the Cell Cycle in Synchronized Cultures of Catharanthus roseus Cells.
Plant Physiol. 1991 Feb;95(2):406-11
PMID: 16667998
-
Nuclear proto-oncogenes fos and jun.
Annu Rev Cell Biol. 1990;6:539-57
PMID: 2125830
-
Neoplastic transformation is associated with coordinate induction of nuclear and cytoplasmic oxidative phosphorylation genes.
J Biol Chem. 1990 Nov 25;265(33):20589-93
PMID: 2173714
-
Molecular cloning of the CD2 antigen, the T-cell erythrocyte receptor, by a rapid immunoselection procedure.
Proc Natl Acad Sci U S A. 1987 May;84(10):3365-9
PMID: 2437578
-
The leucine repeat motif in Fos protein mediates complex formation with Jun/AP-1 and is required for transformation.
Cell. 1989 Feb 10;56(3):507-16
PMID: 2492453
-
Sequence requirements for synthetic peptide-mediated translocation to the nucleus.
Mol Cell Biol. 1989 Jun;9(6):2487-92
PMID: 2668735
-
High efficiency transformation of E. coli by high voltage electroporation.
Nucleic Acids Res. 1988 Jul 11;16(13):6127-45
PMID: 3041370
-
Suppression of the neoplastic phenotype by replacement of the RB gene in human cancer cells.
Science. 1988 Dec 16;242(4885):1563-6
PMID: 3201247
-
Characterization of two genes required for the position-effect control of yeast mating-type genes.
EMBO J. 1984 Dec 1;3(12):2817-23
PMID: 6098447
-
A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.
Anal Biochem. 1983 Jul 1;132(1):6-13
PMID: 6312838
-
c-fos protein can induce cellular transformation: a novel mechanism of activation of a cellular oncogene.
Cell. 1984 Jan;36(1):51-60
PMID: 6607118
-
3' non-coding region sequences in eukaryotic messenger RNA.
Nature. 1976 Sep 16;263(5574):211-4
PMID: 822353
-
A universal method for the direct cloning of PCR amplified nucleic acid.
Biotechnology (N Y). 1991 Jul;9(7):657-63
PMID: 1367662
-
A rapid and efficient protocol for sequencing plasmid DNA.
Biotechniques. 1992 Feb;12(2):228, 230
PMID: 1616715
-
IP-1: a dominant inhibitor of Fos/Jun whose activity is modulated by phosphorylation.
Cell. 1991 Mar 8;64(5):983-93
PMID: 1900458
-
Mitochondrial protein import: isolation and characterization of the Saccharomyces cerevisiae MFT1 gene.
Mol Gen Genet. 1991 Mar;225(3):483-91
PMID: 2017143
-
Revertants of v-fos-transformed rat fibroblasts: suppression of transformation is dominant.
Mol Cell Biol. 1990 Nov;10(11):5626-33
PMID: 2172782
-
Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death.
Nature. 1990 Nov 22;348(6299):334-6
PMID: 2250705
-
Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
Anal Biochem. 1987 Apr;162(1):156-9
PMID: 2440339
-
Detection of genes with a potential for suppressing the transformed phenotype associated with activated ras genes.
Proc Natl Acad Sci U S A. 1989 Jan;86(1):162-6
PMID: 2463620
-
A ras-related gene with transformation suppressor activity.
Cell. 1989 Jan 13;56(1):77-84
PMID: 2642744
-
Molecular and biochemical characterization of the human trk proto-oncogene.
Mol Cell Biol. 1989 Jan;9(1):24-33
PMID: 2927393
-
Nonreciprocal exchanges of information between DNA duplexes coinjected into mammalian cell nuclei.
Mol Cell Biol. 1985 Jan;5(1):59-69
PMID: 2984556
-
Partial reversion of the transformed phenotype in HRAS-transfected tumorigenic cells by transfer of a human gene.
Proc Natl Acad Sci U S A. 1988 Mar;85(5):1590-4
PMID: 3125552
-
Revertants of v-fos-transformed fibroblasts have mutations in cellular genes essential for transformation by other oncogenes.
Cell. 1987 Nov 6;51(3):357-69
PMID: 3664639
-
Hygromycin B phosphotransferase as a selectable marker for DNA transfer experiments with higher eucaryotic cells.
Mol Cell Biol. 1984 Dec;4(12):2929-31
PMID: 6098829
-
FBJ murine osteosarcoma virus: identification and molecular cloning of biologically active proviral DNA.
J Virol. 1982 Nov;44(2):674-82
PMID: 6292525
-
Lambda replacement vectors carrying polylinker sequences.
J Mol Biol. 1983 Nov 15;170(4):827-42
PMID: 6315951
-
Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs.
Nucleic Acids Res. 1984 Jan 25;12(2):857-72
PMID: 6694911
-
Transformation effector and suppressor genes.
J Cell Biochem. 1991 Jul;46(3):199-205
PMID: 1774222