Expression of cloned mitochondrial DNA from the petite negative yeast Schizosaccharomyces pombe in E. coli minicells

1983 ◽  
Vol 191 (1) ◽  
pp. 91-98 ◽  
Author(s):  
L. Del Giudice ◽  
K. Wolf ◽  
F. Manna ◽  
D. R. Massardo
Keyword(s):  
Mitochondrial Dna ◽  
Schizosaccharomyces Pombe ◽  
E Coli ◽  
Petite Negative Yeast

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 Nuclear mutations in the petite-negative yeast Schizosaccharomyces pombe allow growth of cells lacking mitochondrial DNA.

Genetics ◽  
1992 ◽  
Vol 131 (2) ◽  
pp. 255-260 ◽  
Author(s):  
P Haffter ◽  
T D Fox
Keyword(s):  
Mitochondrial Dna ◽  
Schizosaccharomyces Pombe ◽  
Ethidium Bromide ◽  
Blot Hybridization ◽  
Potassium Acetate ◽  
Whole Cells ◽  
Nuclear Mutations ◽  
Petite Negative Yeast ◽  
Derivatives Of

Abstract The fission yeast Schizosaccharomyces pombe has never been found to give rise to viable cells totally lacking mitochondrial DNA (rho(o)). This paper describes the isolation of rho(o) strains of S. pombe by very long term incubation of cells in liquid medium containing glucose, potassium acetate and ethidium bromide. Once isolated, the rho(o) strains did not require potassium acetate or any other novel growth factors. These nonrespiring strains contained no mitochondrial DNA (mtDNA) detectable either by gel-blot hybridization using as probe a clone containing the entire S. pombe mtDNA, or by 1',6-diamidino-2-phenylindole staining of whole cells. Induction of rho(o) derivatives of standard laboratory strains was not reproducible from culture to culture. The cause of this irreproducibility appears to be that growth of the rho(o) strains of S. pombe depended on nuclear mutations that occurred in some, but not all, of the initial cultures. Two independent rho(o) isolates contained mutations in unlinked genes, termed ptp1-1 and ptp2-1. These mutations allowed reproducible ethidium bromide induction of viable rho(o) strains. No other phenotypes were associated with ptp mutations in rho+ strains.

Quantitative PCR for detection of DNA damage in mitochondrial DNA of the fission yeast Schizosaccharomyces pombe

2016 ◽  
Vol 127 ◽  
pp. 77-81 ◽  
Author(s):  
Takanori Senoo ◽  
Mayumi Yamanaka ◽  
Atori Nakamura ◽  
Tomoki Terashita ◽  
Shinji Kawano ◽  
...  
Keyword(s):  
Dna Damage ◽  
Mitochondrial Dna ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Quantitative Pcr

scholarly journals Molecular cloning of mtp70, a mitochondrial member of the hsp70 family.

1989 ◽  
Vol 9 (11) ◽  
pp. 5163-5168 ◽  
Author(s):  
D M Engman ◽  
L V Kirchhoff ◽  
J E Donelson
Keyword(s):  
Mitochondrial Dna ◽  
Dna Replication ◽  
Protozoan Parasite ◽  
Specific Region ◽  
Kinetoplast Dna ◽  
Intracellular Location ◽  
E Coli ◽  
Hsp70 Family ◽  
Mitochondrial Dna Replication ◽  
Initiation Of Dna Replication

We have isolated a gene from the protozoan parasite Trypanosoma cruzi that encodes a previously unidentified member of the 70-kilodalton heat shock protein (hsp70) family. Among all the eucaryotic hsp70 proteins described to date, this trypanosome protein, mtp70, is uniquely related in sequence and structure to the hsp70 of Escherichia coli, DnaK, which functions in the initiation of DNA replication. This relationship to DnaK is especially relevant in view of the intracellular location of the protein. Within the trypanosome, mtp70 is located in the mitochondrion, where it associates with kinetoplast DNA (kDNA), the unusual mitochondrial DNA that distinguishes this order of protozoa. Moreover, mtp70 is located in the specific region of the kinetoplast in which kDNA replication occurs. In view of the known functions of DnaK, the localization of mtp70 to the site of kDNA replication suggests that mtp70 may participate in eucaryotic mitochondrial DNA replication in a manner analogous to that of DnaK in E. coli.

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