scholarly journals Effect of heterologous expression of acyl-CoA-binding protein on acyl-CoA level and composition in yeast

1993 ◽  
Vol 290 (2) ◽  
pp. 369-374 ◽  
Author(s):  
S Mandrup ◽  
R Jepsen ◽  
H Skøtt ◽  
J Rosendal ◽  
P Højrup ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Growth Rate ◽  
Heterologous Expression ◽  
Binding Protein ◽  
Cellular Protein ◽  
Serine Residue ◽  
Yeast Saccharomyces Cerevisiae ◽  
Total Cellular Protein ◽  
Gal1 Promoter

We have expressed a bovine synthetic acyl-CoA-binding protein (ACBP) gene in yeast (Saccharomyces cerevisiae) under the control of the GAL1 promoter. The heterologously expressed bovine ACBP constituted up to 6.4% of total cellular protein and the processing was identical with that of native bovine ACBP, i.e. the initiating methionine was removed and the following serine residue was N-acetylated. The expression of this protein did not affect the growth rate of the cells. Determination of the yeast acyl-CoA pool size showed a close positive correlation between the ACBP content of the cells and the size of the acyl-CoA pool. Thus ACBP can act as an intracellular acyl-CoA pool former. Possible physiological functions of ACBP in cells are discussed.

scholarly journals Maintenance of Mitochondrial Morphology Is Linked to Maintenance of the Mitochondrial Genome in Saccharomyces cerevisiae

Genetics ◽  
2002 ◽  
Vol 162 (3) ◽  
pp. 1147-1156 ◽  
Author(s):  
Theodor Hanekamp ◽  
Mary K Thorsness ◽  
Indrani Rebbapragada ◽  
Elizabeth M Fisher ◽  
Corrine Seebart ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Growth Rate ◽  
Mitochondrial Dna ◽  
Carbon Sources ◽  
Mitochondrial Morphology ◽  
Yeast Saccharomyces Cerevisiae ◽  
Slow Growth Rate ◽  
Dna Migration ◽  
Mitochondrial Dna Stability ◽  
Extragenic Suppressors

Abstract In the yeast Saccharomyces cerevisiae, certain mutant alleles of YME4, YME6, and MDM10 cause an increased rate of mitochondrial DNA migration to the nucleus, carbon-source-dependent alterations in mitochondrial morphology, and increased rates of mitochondrial DNA loss. While single mutants grow on media requiring mitochondrial respiration, any pairwise combination of these mutations causes a respiratory-deficient phenotype. This double-mutant phenotype allowed cloning of YME6, which is identical to MMM1 and encodes an outer mitochondrial membrane protein essential for maintaining normal mitochondrial morphology. Yeast strains bearing null mutations of MMM1 have altered mitochondrial morphology and a slow growth rate on all carbon sources and quantitatively lack mitochondrial DNA. Extragenic suppressors of MMM1 deletion mutants partially restore mitochondrial morphology to the wild-type state and have a corresponding increase in growth rate and mitochondrial DNA stability. A dominant suppressor also suppresses the phenotypes caused by a point mutation in MMM1, as well as by specific mutations in YME4 and MDM10.

scholarly journals A glucan from active dry baker’s yeast (Saccharomyces cerevisiae): A chemical and enzymatic investigation of the structure

2003 ◽  
Vol 68 (11) ◽  
pp. 805-809 ◽  
Author(s):  
Dragan Zlatkovic ◽  
Dragica Jakovljevic ◽  
Djordje Zekovic ◽  
Miroslav Vrvic
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Optical Rotation ◽  
Linear Chain ◽  
Periodate Oxidation ◽  
Methylation Analysis ◽  
Yeast Saccharomyces Cerevisiae ◽  
New Approach ◽  
Main Structural Feature ◽  
Or Groups

The structure of a polysaccharide consisting of D-glucose isolated from the cell-wall of active dry baker?s yeast (Saccharomyces cerevisiae) was investigated by using methylation analysis, periodate oxidation, mass spectrometry, NMR spectroscopy, and enzymic hydrolysis, as a new approach in determination of structures. The main structural feature of the polysaccharide deduced on the basis of the obtained results is a linear chain of (1?3)-linked ?-D-glucopyranoses, a part of which is substituted through the positions O-6. The side units or groups are either a single D-glucopyranose or (1?3)-?-oligoglucosides, linked to the main chaing through (1?6)-glucosidic linkages. The low optical rotation as well as the 13C-NMR and FTIR spectra suggest that the glycosidic linkages are in the ?-D-configuration.

mRNA cap-binding protein: cloning of the gene encoding protein synthesis initiation factor eIF-4E from Saccharomyces cerevisiae

1987 ◽  
Vol 7 (3) ◽  
pp. 998-1003
Author(s):  
M Altmann ◽  
C Handschin ◽  
H Trachsel
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Synthesis ◽  
Genomic Library ◽  
Binding Protein ◽  
Initiation Factor ◽  
Cdna Clones ◽  
Yeast Saccharomyces Cerevisiae ◽  
Peptide Pattern ◽  
Protein Synthesis Initiation ◽  
Encoding Protein

We have isolated genomic and cDNA clones encoding protein synthesis initiation factor eIF-4E (mRNA cap-binding protein) of the yeast Saccharomyces cerevisiae. Their identity was established by expression of a cDNA in Escherichia coli. This cDNA encodes a protein indistinguishable from purified eIF-4E in terms of molecular weight, binding to and elution from m7GDP-agarose affinity columns, and proteolytic peptide pattern. The eIF-4E gene was isolated by hybridization of cDNA to clones of a yeast genomic library. The gene lacks introns, is present in one copy per haploid genome, and encodes a protein of 213 amino acid residues. Gene disruption experiments showed that the gene is essential for growth.

scholarly journals The Lipomyces starkeyi gene Ls120451 encodes a cellobiose transporter that enables cellobiose fermentation in Saccharomyces cerevisiae

2020 ◽  
Vol 20 (3) ◽  
Author(s):  
Jorg C de Ruijter ◽  
Kiyohiko Igarashi ◽  
Merja Penttilä
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Aspergillus Niger ◽  
Heterologous Expression ◽  
In Silico ◽  
Penicillium Oxalicum ◽  
Microbial Fermentation ◽  
Lipomyces Starkeyi ◽  
Yeast Saccharomyces Cerevisiae ◽  
Sugar Transporters ◽  
Selection Of

ABSTRACT Processed lignocellulosic biomass is a source of mixed sugars that can be used for microbial fermentation into fuels or higher value products, like chemicals. Previously, the yeast Saccharomyces cerevisiae was engineered to utilize its cellodextrins through the heterologous expression of sugar transporters together with an intracellular expressed β-glucosidase. In this study, we screened a selection of eight (putative) cellodextrin transporters from different yeast and fungal hosts in order to extend the catalogue of available cellobiose transporters for cellobiose fermentation in S. cerevisiae. We confirmed that several in silico predicted cellodextrin transporters from Aspergillus niger were capable of transporting cellobiose with low affinity. In addition, we found a novel cellobiose transporter from the yeast Lipomyces starkeyi, encoded by the gene Ls120451. This transporter allowed efficient growth on cellobiose, while it also grew on glucose and lactose, but not cellotriose nor cellotetraose. We characterized the transporter more in-depth together with the transporter CdtG from Penicillium oxalicum. CdtG showed to be slightly more efficient in cellobiose consumption than Ls120451 at concentrations below 1.0 g/L. Ls120451 was more efficient in cellobiose consumption at higher concentrations and strains expressing this transporter grew slightly slower, but produced up to 30% more ethanol than CdtG.

scholarly journals Role of IME1 expression in regulation of meiosis in Saccharomyces cerevisiae.

1990 ◽  
Vol 10 (12) ◽  
pp. 6103-6113 ◽  
Author(s):  
H E Smith ◽  
S S Su ◽  
L Neigeborn ◽  
S E Driscoll ◽  
A P Mitchell
Keyword(s):  
Gene Expression ◽  
Saccharomyces Cerevisiae ◽  
Amino Acid ◽  
Specific Gene ◽  
Alpha Cells ◽  
Cell Type ◽  
Yeast Saccharomyces Cerevisiae ◽  
Specific Gene Expression ◽  
Gal1 Promoter ◽  
Acid Protein

Two signals are required for meiosis and spore formation in the yeast Saccharomyces cerevisiae: starvation and the MAT products a1 and alpha 2, which determine the a/alpha cell type. These signals lead to increased expression of the IME1 (inducer of meiosis) gene, which is required for sporulation and sporulation-specific gene expression. We report here the sequence of the IME1 gene and the consequences of IME1 expression from the GAL1 promoter. The deduced IME1 product is a 360-amino-acid protein with a tyrosine-rich C-terminal region. Expression of PGAL1-IME1 in vegetative a/alpha cells led to moderate accumulation of four early sporulation-specific transcripts (IME2, SPO11, SPO13, and HOP1); the transcripts accumulated 3- to 10-fold more after starvation. Two sporulation-specific transcripts normally expressed later (SPS1 and SPS2) did not accumulate until PGAL1-IME1 strains were starved, and the intact IME1 gene was not activated by PGAL1-IME1 expression. In a or alpha cells, which lack alpha 2 or a1, expression of PGAL1-IME1 led to the same pattern of IME2 and SPO13 expression as in a/alpha cells, as measured with ime2::lacZ and spo13::lacZ fusions. Thus, in wild-type strains, the increased expression of IME1 in starved a/alpha cells can account entirely for cell type control, but only partially for nutritional control, of early sporulation-specific gene expression. PGAL1-IME1 expression did not cause growing cells to sporulate but permitted efficient sporulation of amino acid-limited cells, which otherwise sporulated poorly. We suggest that IME1 acts primarily as a positive regulator of early sporulation-specific genes and that growth arrest is an independent prerequisite for execution of the sporulation program.

scholarly journals A novel FK506- and rapamycin-binding protein (FPR3 gene product) in the yeast Saccharomyces cerevisiae is a proline rotamase localized to the nucleolus.

1994 ◽  
Vol 127 (3) ◽  
pp. 623-639 ◽  
Author(s):  
B M Benton ◽  
J H Zang ◽  
J Thorner
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Product ◽  
Full Length ◽  
Multicopy Plasmid ◽  
Yeast Saccharomyces Cerevisiae ◽  
Normal Cells ◽  
Terminal Domain ◽  
Growth Inhibitory ◽  
Gal1 Promoter ◽  
Delta Cells

The gene (FPR3) encoding a novel type of peptidylpropyl-cis-trans-isomerase (PPIase) was isolated during a search for previously unidentified nuclear proteins in Saccharomyces cerevisiae. PPIases are thought to act in conjunction with protein chaperones because they accelerate the rate of conformational interconversions around proline residues in polypeptides. The FPR3 gene product (Fpr3) is 413 amino acids long. The 111 COOH-terminal residues of Fpr3 share greater than 40% amino acid identity with a particular class of PPIases, termed FK506-binding proteins (FKBPs) because they are the intracellular receptors for two immunosuppressive compounds, rapamycin and FK506. When expressed in and purified from Escherichia coli, both full-length Fpr3 and its isolated COOH-terminal domain exhibit readily detectable PPIase activity. Both fpr3 delta null mutants and cells expressing FPR3 from its own promoter on a multicopy plasmid have no discernible growth phenotype and do not display any alteration in sensitivity to the growth-inhibitory effects of either FK506 or rapamycin. In S. cerevisiae, the gene for a 112-residue cytosolic FKBP (FPR1) and the gene for a 135-residue ER-associated FKBP (FPR2) have been described before. Even fpr1 fpr2 fpr3 triple mutants are viable. However, in cells carrying an fpr1 delta mutation (which confers resistance to rapamycin), overexpression from the GAL1 promoter of the C-terminal domain of Fpr3, but not full-length Fpr3, restored sensitivity to rapamycin. Conversely, overproduction from the GAL1 promoter of full-length Fpr3, but not its COOH-terminal domain, is growth inhibitory in both normal cells and fpr1 delta mutants. In fpr1 delta cells, the toxic effect of Fpr3 overproduction can be reversed by rapamycin. Overproduction of the NH2-terminal domain of Fpr3 is also growth inhibitory in normal cells and fpr1 delta mutants, but this toxicity is not ameliorated in fpr1 delta cells by rapamycin. The NH2-terminal domain of Fpr3 contains long stretches of acidic residues alternating with blocks of basic residues, a structure that resembles sequences found in nucleolar proteins, including S. cerevisiae NSR1 and mammalian nucleolin. Indirect immunofluorescence with polyclonal antibodies raised against either the NH2- or the COOH-terminal segments of Fpr3 expressed in E. coli demonstrated that Fpr3 is located exclusively in the nucleolus.

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