Abstract
We identified and isolated a Saccharomyces cerevisiae gene which, when overexpressed, suppressed the temperature-sensitive phenotype of cells expressing a mutant allele of the gene encoding the mitochondrial chaperonin, Hsp60. This gene, SCS1 (suppressor of chaperonin sixty-1), encodes a 757-amino-acid protein of as yet unknown function which, nonetheless, has human, rice, and Caenorhabditis elegans homologs with high degrees (ca. 60%) of amino acid sequence identity. SCS1 is not an essential gene, but SCS1-null strains do not grow above 37 degrees C and show some growth-related defects at 30 degrees C as well. This gene is expressed at both 30 and 38 degrees C, producing little or no differences in mRNA levels at these two temperatures. Overexpression of SCS1 could not complement an HSP60-null allele, indicating that suppression was not due to the bypassing of Hsp60 activity. Of 10 other hsp60-ts alleles tested, five could also be suppressed by SCS1 overexpression. There were no common mutant phenotypes of the strains expressing these alleles that give any clue as to why they were suppressible while others were not. An epitope (influenza virus hemagglutinin)-tagged form of SCS1 in single copy complemented an SCS1-null allele. The Scs1-hemagglutinin protein was found to be at comparable levels and in similar multiply modified forms in cells growing at both 30 and 38 degrees C. Surprisingly, when localized either by cell fractionation procedures or by immunocytochemistry, these proteins were found not in mitochondria but in the cytosol. The overexpression of SCS1 had significant effects on the cellular levels of mRNAs encoding the proteins Cpn10 and Mgel, two other mitochondrial protein cochaperones, but not on mRNAs encoding a number of other mitochondrial or cytosolic proteins analyzed. The implications of these findings are discussed.
MeSH Terms
Alleles
Amino Acid Sequence
Base Sequence
Basic Helix-Loop-Helix Transcription Factors
Chaperonin 60/genetics
Cloning, Molecular
Cytosol/chemistry
Fungal Proteins/biosynthesis,chemistry,genetics,physiology
Gene Dosage
Gene Expression Regulation, Fungal/genetics
Genes, Fungal/genetics,physiology
Genes, Suppressor/physiology
Mitochondria/chemistry
Molecular Sequence Data
RNA, Messenger/biosynthesis
Recombinant Fusion Proteins/biosynthesis
Repressor Proteins/biosynthesis,chemistry,genetics,physiology
Saccharomyces cerevisiae Proteins
Sequence Homology, Amino Acid
Temperature
Transcription Factors
Chemicals
Basic Helix-Loop-Helix Transcription Factors
Chaperonin 60
Fungal Proteins
INO2 protein, S cerevisiae
RNA, Messenger
Recombinant Fusion Proteins
Repressor Proteins
Saccharomyces cerevisiae Proteins
Transcription Factors
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Shu Y
Department of Biology, Syracuse University, New York 13244, USA.
Hallberg R L
References (33)
33 references, click to expand
-
Rapid and efficient site-specific mutagenesis without phenotypic selection.
Methods Enzymol. 1987;154:367-82
PMID: 3323813
-
KAR1, a gene required for function of both intranuclear and extranuclear microtubules in yeast.
Cell. 1987 Mar 27;48(6):1047-60
PMID: 3030557
-
Yeast/E. coli shuttle vectors with multiple unique restriction sites.
Yeast. 1986 Sep;2(3):163-7
PMID: 3333305
-
Isolation and characterization of conditional-lethal mutations in the TUB1 alpha-tubulin gene of the yeast Saccharomyces cerevisiae.
Genetics. 1988 Nov;120(3):681-95
PMID: 3066684
-
Mitochondrial heat-shock protein hsp60 is essential for assembly of proteins imported into yeast mitochondria.
Nature. 1989 Feb 16;337(6208):620-5
PMID: 2645524
-
Characterization of the yeast HSP60 gene coding for a mitochondrial assembly factor.
Nature. 1989 Feb 16;337(6208):655-9
PMID: 2563898
-
A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.
Genetics. 1989 May;122(1):19-27
PMID: 2659436
-
SSC1, an essential member of the yeast HSP70 multigene family, encodes a mitochondrial protein.
Mol Cell Biol. 1989 Jul;9(7):3000-8
PMID: 2674677
-
Protein folding in mitochondria requires complex formation with hsp60 and ATP hydrolysis.
Nature. 1989 Sep 14;341(6238):125-30
PMID: 2528694
-
The NUP1 gene encodes an essential component of the yeast nuclear pore complex.
Cell. 1990 Jun 15;61(6):965-78
PMID: 2190694
-
Role of vacuolar acidification in protein sorting and zymogen activation: a genetic analysis of the yeast vacuolar proton-translocating ATPase.
Mol Cell Biol. 1990 Jul;10(7):3737-49
PMID: 2141385
-
Meiotic gene conversion and crossing over: their relationship to each other and to chromosome synapsis and segregation.
Cell. 1990 Sep 7;62(5):927-37
PMID: 2203538
-
Preparation of high molecular weight RNA.
Methods Enzymol. 1991;194:398-405
PMID: 1706459
-
Molecular chaperones.
Annu Rev Biochem. 1991;60:321-47
PMID: 1679318
-
Sequential action of mitochondrial chaperones in protein import into the matrix.
EMBO J. 1991 Nov;10(11):3273-80
PMID: 1915294
-
The ROX3 gene encodes an essential nuclear protein involved in CYC7 gene expression in Saccharomyces cerevisiae.
Mol Cell Biol. 1991 Nov;11(11):5639-47
PMID: 1656237
-
Heat shock and stationary phase induce transcription of the Saccharomyces cerevisiae iso-2 cytochrome c gene.
Curr Genet. 1991 Aug;20(3):185-8
PMID: 1657416
-
Successive action of DnaK, DnaJ and GroEL along the pathway of chaperone-mediated protein folding.
Nature. 1992 Apr 23;356(6371):683-9
PMID: 1349157
-
Import of proteins into mitochondria. Cytochrome b2 and cytochrome c peroxidase are located in the intermembrane space of yeast mitochondria.
J Biol Chem. 1982 Nov 10;257(21):13028-33
PMID: 6290489
-
One-step gene disruption in yeast.
Methods Enzymol. 1983;101:202-11
PMID: 6310324
-
hsp70: nuclear concentration during environmental stress and cytoplasmic storage during recovery.
Cell. 1984 Mar;36(3):655-62
PMID: 6421488
-
Assembly of the mitochondrial membrane system. Characterization of COR1, the structural gene for the 44-kilodalton core protein of yeast coenzyme QH2-cytochrome c reductase.
J Biol Chem. 1986 Dec 25;261(36):17163-9
PMID: 3023384
-
Folding in vivo of bacterial cytoplasmic proteins: role of GroEL.
Cell. 1993 Sep 10;74(5):909-17
PMID: 8104102
-
A highly evolutionarily conserved mitochondrial protein is structurally related to the protein encoded by the Escherichia coli groEL gene.
Mol Cell Biol. 1988 Jan;8(1):371-80
PMID: 2892128
-
Cytochromes c1 and b2 are sorted to the intermembrane space of yeast mitochondria by a stop-transfer mechanism.
Cell. 1992 May 29;69(5):809-22
PMID: 1350514
-
Loss of mitochondrial hsp60 function: nonequivalent effects on matrix-targeted and intermembrane-targeted proteins.
Mol Cell Biol. 1993 May;13(5):3050-7
PMID: 8097278
-
Identification and functional analysis of chaperonin 10, the groES homolog from yeast mitochondria.
Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):10967-71
PMID: 7902576
-
Mdj1p, a novel chaperone of the DnaJ family, is involved in mitochondrial biogenesis and protein folding.
Cell. 1994 Apr 22;77(2):249-59
PMID: 8168133
-
A mitochondrial homolog of bacterial GrpE interacts with mitochondrial hsp70 and is essential for viability.
EMBO J. 1994 Apr 15;13(8):1998-2006
PMID: 8168496
-
A role for a eukaryotic GrpE-related protein, Mge1p, in protein translocation.
Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6481-5
PMID: 8022808
-
Role of the chaperonin cofactor Hsp10 in protein folding and sorting in yeast mitochondria.
J Cell Biol. 1994 Jul;126(2):305-15
PMID: 7913473
-
The crystal structure of the bacterial chaperonin GroEL at 2.8 A.
Nature. 1994 Oct 13;371(6498):578-86
PMID: 7935790
-
Homologous plant and bacterial proteins chaperone oligomeric protein assembly.
Nature. 1988 May 26;333(6171):330-4
PMID: 2897629