Interactions between the yeast mitochondrial and nuclear genomes: isogenic suppressive and hypersuppressive petites differ in their resistance to the alkaloid lycorine

In a previous paper we have shown that the alkaloid lycorine inhibits growth of rho+, mit- and rho-, strains of Saccharomyces cerevisiae, whereas strains devoid of mitochondrial DNA (rho degrees) are resistant to more than 200 micrograms/ml of the alkaloid. In this report we show that hypersuppressive petites are almost as resistant as rho degrees mutants, whereas isogenic rho- petites, which have retained longer segments of the genome, are sensitive to the drug.

Nucleotide sequence of the unassigned reading frame urf a in the mitochondrial genome of three Schizosaccharomyces pombe strains.

Scanning electron microscopy of heterochromatin in chromosome spreads of male germ cells in Schistocerca gregaria (Acrididae, Orthoptera) after trypsinization.

Chromosome spreads, prepared from testes of the desert locust Schistocerca gregaria, were analyzed using scanning electron microscopy (SEM) after varying periods of preincubation in trypsin. The emphasis of the study was on the appearance of heterochromatin. A trypsin pretreatment of 5 sec resulted in a smooth surface on the chromatin throughout and the heterochromatin was highly electron-emissive. The facultatively heterochromatic X chromosome was clearly visible in interphase spermatogonia and in pachytene and late prophase I spermatocytes. Chromomeres of autosomal bivalents could be recognized in pachytene cells. Centromeric heterochromatin segments were very prominent in autosomes of la…

Centromere-Like Elements in Megaselia Spiracularis (Diptera: Phoridae): A Fine-Structure and Cytogenetic Study

The present study was concerned with the karyotype of a humpbacked fly, Megaselia spiracularis (Diptera: Phoridae). Chromosome preparations of neuroblasts isolated from male and female pupae uniformly showed two pairs of metacentric chromosomes and a telocentric chromosome pair. All chromosomes were homomorphic. The analysis of ultrathin serial sections through spermatogonia in metaphase using transmission electron microscopy, confirmed the presence of 3 regular chromosome pairs. In ultrathin sections, the centromeres could be detected as individual elements owing to their lesser electron density in comparison with the chromosome arms. In addition, the spindles contained two tiny elements s…

The pattern of histone H4 acetylation on the X chromosome during spermatogenesis of the desert locustSchistocerca gregaria

We have used antibodies directed against histone H4 acetylated at lysine residue 5, 8, 12, or 16 and indirect immunofluorescence microscopy to probe chromosomes from spermatogonia and spermatocytes of the desert locust, Schistocerca gregaria. The autosomes showed bright overall fluorescence, indicative of high levels of H4 acetylation. In contrast, the X chromosome, which is facultatively heterochromatic during spermatogenesis of the locust, remained completely unstained in spermatogonia and secondary spermatocytes and showed only a small terminal fluorescent band in primary spermatocytes. This band probably corresponds to centromere-associated constitutive heterochromatin. Thus, underacety…

Strange vesicles with a homogeneous content in spermatocytes and spermatids of a click beetle, Adelocera murina (Elateridae). A fine structure study

Abstract The restructuring of primary spermatocytes of Adelocera murina, a click beetle, is described using electron microscopy of ultrathin sections. Emphasis is on spherical or rod-shaped cytoplasmic inclusions, invested by a unit membrane. The content of the inclusions is slightly more electron-dense than that of the surrounding cytoplasm and homogeneously textured in most cases. The inclusions are missing in spermatogonia but are abundant in prophase I through anaphase I spermatocytes. Their number declines in telophase I. Very similar elements are associated with the distal ends of the outgrowing flagella in metaphase I through telophase I spermatocytes and form the so-called flagellar…

Characterization of a novel open reading frame, urf a, in the mitochondrial genome of fission yeast: correlation of urf a mutations with a mitochondrial mutator phenotype and a possible role of frameshifting in urf a expression.

Between the genes for tRNA(gin) and tRNA(ile) an open reading frame of 227 amino acids has been identified which is unique among known mitochondrial genomes and which has been termed urf a (Lang et al. 1983; Kornrumpf et al. 1984). It uses the "mitochondrial" genetic code, i.e., it contains a TGA codon, whereas all other protein-encoding genes, and all but one intronic open reading frame, use the "standard" genetic code (UGG for tryptophan). A previous paper has demonstrated that "mutator" strains show an increased formation of mitochondrial drug-resistant and respiration-deficient mutants (including deletions). In this paper we show that the mutator activity is correlated with mutations in…

The mitochondrial genome of fission yeast: inability of all introns to splice autocatalytically, and construction and characterization of an intronless genome

In this paper we report the inability of four group I introns in the gene encoding subunit I of cytochrome c oxidase (cox1) and the group II intron in the apocytochrome b gene (cob) to splice autocatalytically. Furthermore we present the characterization of the first cox1 intron in the mutator strain anar-14 and the construction and characterization of strains with intronless mitochondrial genomes. We provide evidence that removal of introns at the DNA level (termed DNA splicing) is dependent on an active RNA maturase. Finally we demonstrate that the absence of introns does not abolish homologous mitochondrial recombination.

The mitochondrial genome of Schizosaccharomyces pombe. Stimulation of intra-chromosomal recombination in Escherichia coli by the gene product of the first cox1 intron.

The open reading frame of the first intron of the mitochondrial cox1 gene (cox1I1) was expressed in Escherichia coli. The putative intron-encoded protein stimulated the formation of intra-chromosomal lac +-recombinants about threefold. No stimulation was found when the reading frame was inserted in the opposite direction, or when it was interrupted by a deletion. The intronic open reading frame did not complement recA − or recB − mutants of E. coli. In S. pombe, elimination of this intron did not abolish homologous recombination in mitochondria. A possible role of the recombinase activity in yeast mitochondria will be discussed.

Treatment with the anti-tumor drugs, cis-platin and mafosfamide, does not affect the structure of prekinetochores in a human breast cancer cell line. An immunofluorescence study using human anticentromere autoantibodies

Abstract The goal of the present article was to determine whether a nuclear parameter, centromere structure of interphase cells, could serve as an indicator to assess cellular damage caused by anti-tumor drugs. These were cis-platin and mafosfamide, which are widely used for the management of solid tumors. To visualize the centromeres, we probed treated and untreated cells of a human breast cancer cell line, MX-1, with a human anti-centromere serum. The serum was obtained from a scleroderma patient and detects antigens associated with prekinetochores of the decondensed chromosomes. The DNA was simultaneously displayed by a specific fluorescent dye. The cells were grown on coverslips, incuba…

Microtubule organization and distribution of gamma-tubulin in male meiosis of lepidoptera

Meiotic spindles in males of higher Lepidotera are unusual in that the bulk of the spindle micro-tubules (MTs) ends about halfway between the equatorial plate and the centrosomes in metaphase. It appears worthwhile to determine how the MTs are nucleated, while their pole proximal ends are distant from the centrosomes. To this end, spermatocytes of Phragmatobia fuliginosa (Arctiidae), collected in the field, were double-labeled with antibodies to beta- and gamma-tubulin. The former antibody reveals the entire microtubular cytoskeleton, and the latter is directed against a newly-discovered tublin isoform that is prevalent in microtubule-organizing centers (MTOCs). The immunocytochemical work …

Virus-like particles, bacteria and microsporidia affect spindle-associated membranes in spermatocytes of Lepidoptera species.

SummaryLarval testes of four Lepidoptera species were examined using electron microscopy. The testes of one species, the Mediterranean mealmothEphestia kuehniella(Pyralidae), were devoid of intracellular pathogens and serve as a control. In this species, metaphase spindles of primary spermatocytes showed a thick layer of perispindle membranes. The membranes were structurally very similar to the agranular endoplasmic reticulum. Membranes of this type occurred also at high frequency throughout the spindle matrix. The analysis of larval testes ofPieris brassicae(Pieridae) revealed virus-like particles within spermatocytes. In another species,Philudoria potatoria(Lasiocampidae), the spermatocyt…

Organisation von Mikrotubuli in der Zelle

Mikrotubuli (MT) sind obligate Strukturelemente eukaryotischer Zellen. Zusammen mit Mikrofilamenten und intermediaren Filamenten bilden die MT ein cytoplasmatisches Netzwerk, das Cytoskelett. Die fadenformigen Mikrofilamente (Durchmesser etwa 6 nm) und die intermediaren Filamente (Durchmesser etwa 10 nm) sind wesentlich dunner als die MT (Durchmesser etwa 24 nm), die sich durch einen rohrenformigen Aufbau auszeichnen. MT bestehen aus zwei verschiedenen Proteinklassen: Die Tubuline stellen die Hauptkomponente der MT dar und sind obligat vorhanden. In Abhangigkeit vom Zelltyp und der Funktion sind mit den MT weitere Proteine verknupft, die MT-assoziierten Proteine (MAPs). MT liegen in aller R…

Centrosome structure is very similar in eupyrene and apyrene spermatocytes ofEphestia kuehniella(Pyralidae, Lepidoptera, Insecta)

Summary The higher Lepidoptera produce two types of sperm, eupyrene and apyrene. Eupyrene spermatozoa are fertile, while apyrene spermatozoa are anucleate and, therefore, sterile. Meiosis differs between the two developmental lines. One of the most obvious differences is the aberrant organization of the apyrene spindles. They possess fewer microtubules than eupyrene spindles and chromosome segregation is irregular. To determine whether micotubule nucleation is impaired in the aprene spindles, the present fine structure study concentrated on the centrosomes in both spermatocyte types. In addition, the presence of gamma-tubulin, a newly discovered tubulin isoform which is prevalent in centros…

Optical illusions in scanning electron micrographs: the case of the eggshell of Acrosternum (Chinavia) marginatum (Hemiptera: Pentatomidae)

Scanning electron microscopy revealed that-as is common in this family of the Hemiptera-the eggs of the green stink bug Acrosternum (Chinavia) marginatum are roughly barrel-shaped and possess at their apical pole a row of slender extensions, the aero-micropylar processes. The outer surface of the eggshell carries hexagonally arranged pits. The analysis of cross-fractured eggshells showed that the pits have slender basal extensions with transverse diaphragms. When scanning electron micrographs of the egg surface of A. marginatum are viewed upside down, the perception flips and the pits appear as elevations to all observers addressed. Thus, we are dealing with an optical illusion, which is kn…

The Mu1 transposable element of maize contains two promoter signals recognized by the Escherichia coli RNA polymerase.

The galactokinase (GalK) expression plasmid vector system pKO-1 has been used to screen for promoter elements in the maize transposable element Mu1 that function in Escherichia coli. Two transcriptional start points, named S1 and S2, were identified, which are located in the two direct repeats of the transposable element. This paper demonstrates that sequence elements exist in a plant transposable element which function as prokaryotic promotors.

Glutathione metabolism and heavy metal detoxification in Schizosaccharomyces pombe

Sixty glutathione-deficient mutants (gsh −) of Schizosaccharomyces pombe have been isolated by their resistance towards the mutagen N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and their sensitivity to the heavy metal Cadmium (Cd). fifty-three mutants show glutathione contents of less than 5% compared with the wild-type. The residual glutathione contents correlate with the resistance to MNNG, with the sensitivity to Cd and with the growth rate in minimal medium. The gsh −, Cd-sensitive (Cd s) mutants also show sensitivity to other heavy metals. Wild-type strains, but not the gsh − mutants, are able to excrete the heavy metal, very likely as a sulfide-containing compound. This inability of th…

Distribution of gamma‐tubulin differs in primary and secondary oocytes of Ephestia kuehniella (Pyralidae, Lepidoptera)

In a previous study, barrel-shaped spindles were found in metaphase I oocytes of Ephestia kuehniella (Pyralidae, Lepidoptera). Aster microtubules (MTs) were missing (Wolf, 1993 : Cell Motil Cytoskeleton 24 :200-204). This points to an acentriolar organization of the spindle apparatus. The present study was aimed at the question of whether gamma-tubulin, a newly detected member of the tubulin superfamily that has often been identified in microtubule-organizing centers, plays a role in the nucleation of MTs in meiotic spindles of the moth. To this end, the distribution of gamma-tubulin was examined in oocytes of E. kuehniella using an antibody against gamma-tubulin in combination with indirec…