Determination of chromosome copy numbers in Saccharomyces cerevisiae strains via integrative probe and blot hybridization techniques

1995 ◽  
Vol 27 (3) ◽  
pp. 217-228 ◽  
Author(s):  
Christopher Hadfield ◽  
Jennifer A. Harikrishna ◽  
Jacqueline A. Wilson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Blot Hybridization ◽  
Copy Numbers

scholarly journals Expression of the Saccharomyces cerevisiae Gene YME1 in the Petite-Negative Yeast Schizosaccharomyces pombe Converts It to Petite-Positive

Genetics ◽  
2000 ◽  
Vol 154 (1) ◽  
pp. 147-154 ◽  
Author(s):  
Douglas J Kominsky ◽  
Peter E Thorsness
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mitochondrial Dna ◽  
Schizosaccharomyces Pombe ◽  
Atp Synthase ◽  
Kluyveromyces Lactis ◽  
Blot Hybridization ◽  
Inner Mitochondrial Membrane ◽  
Yeast Saccharomyces Cerevisiae ◽  
Mitochondrial Atp Synthase ◽  
Petite Negative Yeast

Abstract Organisms that can grow without mitochondrial DNA are referred to as “petite-positive” and those that are inviable in the absence of mitochondrial DNA are termed “petite-negative.” The petite-positive yeast Saccharomyces cerevisiae can be converted to a petite-negative yeast by inactivation of Yme1p, an ATP- and metal-dependent protease associated with the inner mitochondrial membrane. Suppression of this yme1 phenotype can occur by virtue of dominant mutations in the α- and γ-subunits of mitochondrial ATP synthase. These mutations are similar or identical to those occurring in the same subunits of the same enzyme that converts the petite-negative yeast Kluyveromyces lactis to petite-positive. Expression of YME1 in the petite-negative yeast Schizosaccharomyces pombe converts this yeast to petite-positive. No sequence closely related to YME1 was found by DNA-blot hybridization to S. pombe or K. lactis genomic DNA, and no antigenically related proteins were found in mitochondrial extracts of S. pombe probed with antisera directed against Yme1p. Mutations that block the formation of the F1 component of mitochondrial ATP synthase are also petite-negative. Thus, the F1 complex has an essential activity in cells lacking mitochondrial DNA and Yme1p can mediate that activity, even in heterologous systems.

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.

Isolation of the SUP45 omnipotent suppressor gene of Saccharomyces cerevisiae and characterization of its gene product

1985 ◽  
Vol 5 (4) ◽  
pp. 816-822
Author(s):  
H J Himmelfarb ◽  
E Maicas ◽  
J D Friesen
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ribosomal Protein ◽  
Blot Hybridization ◽  
Single Copy ◽  
Suppressor Gene ◽  
Yeast Cells ◽  
In Vitro Translation ◽  
Nonsense Suppression ◽  
Synthesis Rate

The Saccharomyces cerevisiae SUP45+ gene has been isolated from a genomic clone library by genetic complementation of paromomycin sensitivity, which is a property of a mutant strain carrying the sup45-2 allele. This plasmid complements all phenotypes associated with the sup45-2 mutation, including nonsense suppression, temperature sensitivity, osmotic sensitivity, and paromomycin sensitivity. Genetic mapping with a URA3+-marked derivative of the complementing plasmid that was integrated into the chromosome by homologous recombination demonstrated that the complementing fragment contained the SUP45+ gene and not an unlinked suppressor. The SUP45+ gene is present as a single copy in the haploid genome and is essential for viability. In vitro translation of the hybrid-selected SUP45+ transcript yielded a protein of Mr = 54,000, which is larger than any known ribosomal protein. RNA blot hybridization analysis showed that the steady-state level of the SUP45+ transcript is less than 10% of that for ribosomal protein L3 or rp59 transcripts. When yeast cells are subjected to a mild heat shock, the synthesis rate of the SUP45+ transcript was transiently reduced, approximately in parallel with ribosomal protein transcripts. Our data suggest that the SUP45+ gene does not encode a ribosomal protein. We speculate that it codes for a translation-related function whose precise nature is not yet known.

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