Biochemical characteristics of yeast respiration-deficient mutants differing in buoyant densities of mitochondrial DNA

Zusammenfassung Es wurde nachgewiesen, dass, obwohl die mitochondriale DNS verschiedener, atmungsdefekter Hefemutanten sowohl untereinander als auch von den Wildstämmen unterschieden werden können, diese uniforme biochemische Charakteristika aufweisen: Fehlen von spektroskopisch nachweisbarem Zytochroma.a ₃,b und c₁ gegen Oligomyzin unempfindliche ATPase und die Unfähigkeit, Aminosäuren in Proteine zu inkorporieren.     

A turbidimetric method for the screening of amylase-producing mutants of Aspergillus niger

Summary The method is based on the assumption that extracellular amylase, which is produced by strains ofAspergillus niger in liquid culture, hydrolyses the starch in the media and brings about a corresponding decrease in the turbidity of the media. Mutant strains which produced different quantities of amylase exhibited different degrees of decrease in turbidity of the media. The results showed that a greater degree of decrease in turbidity was observed for a higher quantity of amylase produced.     

Temperature sensitivity in the respiratory-deficient mutants of yeast (Saccharomyces cerevisiae)

Saccharomyces cerevisiae . , .     

Nucleotide sequence of the gene for the mitochondrial 15S ribosomal RNA of yeast

We have determined the nucleotide sequence of a DNA segment carrying the entire 15S ribosomal RNA gene of yeast mitochondrial genome. Many stretches of sequence are present which are homologous to the E. coli 16S ribosomal RNA gene. The gene sequence can be folded into a secondary structure according to the [1] model on bacterial ribosomal RNAs. The structure reveals a striking similarity between the two RNAs despite the large difference in their base compositions. In the middle of the gene, we found a guanine-cytosine rich sequence that is also present in several other regions of the mitochondrial genome.     

ATP-dependent proteases controlling mitochondrial function in the yeast Saccharomyces cerevisiae

Regulated protein degradation by ATP-dependent proteases plays a fundamental role in the biogenesis of mitochondria. Membrane-bound and soluble ATP-dependent proteases have been identified in various subcompartments of this organelle. Subunits composing these proteases are evolutionarily conserved from yeast to humans and, in support of an endosymbiotic origin of mitochondria, evolved from prokaryotic ancestors: the PIM1/Lon protease is active in the matrix of mitochondria, while the i-AAA protease and the m-AAA protease mediate the turnover of inner membrane proteins. Most of the knowledge concerning the biogenesis and the physiological role of ATP-dependent proteases comes from studies in the yeast Saccharomyces cerevisiae. Proteases were found to be required for mitochondrial stasis, for the maintenance of the morphology of the organelle and for mitochondrial genome integrity. ATP-dependent proteolysis is crucial for the expression of mitochondrially encoded subunits of respiratory chain complexes and for the assembly of these complexes. Hence, mitochondrial ATP-dependent proteases exert multiple roles which are essential for the maintenance of cellular respiratory competence.     

Nucleotide sequence determination of yeast mitochondrial phenylalanine-tRNA]

The primary structure of mitochondrial tRNAPhe from Saccharomyces cerevisiae, purified by two-dimensional polyacrylamide gel electrophoresis, was determined using, standard procedures on in vivo 32P-labeled tRNA, as well as the new 5'-end postlabeling techniques. We propose a cloverleaf model which allows for tertiary interaction between cytosine in position 46 and guanine in position 15 and maximizes base pairing in the psi C stem, thus excluding the uracile in position 50 from base pairing in the psi C stem. Comparison of the primary structure of this tRNA with all other known procaryotic, chloroplastic or cytoplasmic tRNAsPhe sequences does not lead to any conclusion about the endosymbiotic theory of mitochondria evolution.     

A new evidence for induction of respiration deficiency in yeast by acriflavine

Zusammenfassung Mit Hilfe einer Nährlösung, in der 2prozentiges Glyzerin den Zucker ersetzte, wurde ein neues und vereinfachendes Verfahren gefunden, die durch Acriflavin bedingte Induktion der Atmungsunfähigkeit von Hefe zu prüfen. Während die normalen, das heisst atmungsfähigen Zellen sich in dieser Nährlösung vermehren, tun dies die atmungsunfähigen Zellen nie. Eine bemerkenswerte Zunahme der atmungsunfähigen Zellen wurde bei normalen Hefekulturen in Glyzerin-Nährlösung nach Zusatz von verschiedenen Konzentrationen von Acriflavin trotz der nachteiligen Bedingungen beobachtet.     

Molecular genetics of the peptidyl transferase center and the unusual Var1 protein in yeast mitochondrial ribosomes

Mitochondria posses their own ribosomes responsible for the synthesis of a small number of proteins encoded by the mitochondrial genome. In yeast,Saccharomyces cerevisiae, the two ribosomal RNAs and a single ribosomal protein, Varl, are products of mitochondrial genes, and the remaining approximately 80 ribosomal proteins are encoded in the nucleus. The mitochondrial translation system is dispensable in yeast, providing an excellent experimental model for the molecular genetic analysis of the fundamental properties of ribosomes in general as well as adaptations required for the specialized role of ribosomes in mitochondria. Recent studies of the peptidyl transferase center, one of the most highly conserved functional centers of the ribosome, and the Varl protein, an unusual yet essential protein in the small ribosomal subunit, have provided new insight into conserved and divergent features of the mitochondrial ribosome.     

The effect of two new peptide antibiotics,the hypelcins,on mitochondrial function

Summary The action on mitochondria of 3 peptide antibiotics, hypelcin-A, hypelcin-B, and alamethicin, was examined. The results showed that they are unique uncouplers of oxidative phosphorylation, with the same mechanism of action.     

Lack of amino acid incorporation by isolated mitochondria from respiratory-deficient cytoplasmic yeast mutants

Zusammenfassung Die Mutation der mitochondrialen DNA, die in den atmungsdefekten Hefen resultiert, führt zu dem Verlust der Fähigkeit der isolierten Mutantenmitochondrien,¹⁴C-Leuzin in die TES-unlösbare Fraktion zu inkorporieren.     

Regulation of mitochondrial dynamics: convergences and divergences between yeast and vertebrates

In eukaryotic cells, the shape of mitochondria can be tuned to various physiological conditions by a balance of fusion and fission processes termed mitochondrial dynamics. Mitochondrial dynamics controls not only the morphology but also the function of mitochondria, and therefore is crucial in many aspects of a cell’s life. Consequently, dysfunction of mitochondrial dynamics has been implicated in a variety of human diseases including cancer. Several proteins important for mitochondrial fusion and fission have been discovered over the past decade. However, there is emerging evidence that there are as yet unidentified proteins important for these processes and that the fusion/fission machinery is not completely conserved between yeast and vertebrates. The recent characterization of several mammalian proteins important for the process that were not conserved in yeast, may indicate that the molecular mechanisms regulating and controlling the morphology and function of mitochondria are more elaborate and complex in vertebrates. This difference could possibly be a consequence of different needs in the different cell types of multicellular organisms. Here, we review recent advances in the field of mitochondrial dynamics. We highlight and discuss the mechanisms regulating recruitment of cytosolic Drp1 to the mitochondrial outer membrane by Fis1, Mff, and MIEF1 in mammals and the divergences in regulation of mitochondrial dynamics between yeast and vertebrates.