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D Zickler

Publications and source records attributed to D Zickler.

29 records · Page 2Linked to original sources

How the cytoskeleton recognizes and sorts nuclei of opposite mating type during the sexual cycle in filamentous ascomycetes.

In heterothallic filamentous ascomycetes, two nuclei of opposite mating type must recognize one another in a plurinucleate cell to form a pair prior to karyogamy. In pseudohomothallic species, two nuclei of opposite mating type must also pair after meiosis to form a binucleate spore. We have examined the cytoskeletal involvement in nuclear pairings by immunofluorescence and drug disruption, using heterothallic and pseudohomothallic species, as well as species without defined mating type (homothallic). Nuclei of species with defined mating type have spindle pole bodies which react with chromatin stains; those of homothallic species do not. The reactivity is seen only in interphase, not during nuclear divisions; thus, the DNA concerned is nuclear and not organellar. From light and immunofluorescence microscopy, the DNA is located at the nuclear face of the spindle pole body (SPB). We suggest that the DNA-SPB association may be involved in the recognition of self and nonself between nuclei of opposite mating types. Nuclei which cooperate in cell formation during ascus development or sporulation are placed in close proximity by the arrangement of spindles during the division preceding cell formation; after division, each nuclear pair remains linked by intertwined microtubule asters. Nuclear pairs must migrate before binucleate spore formation. Drug disruptions established that actin-myosin interaction was the most important cytoskeletal factor in normal spore production. The ascomycete SPB shows unexpected flexibility in form and location during development. Prior to sporulation the outer plaque shows extensive modification in size and orientation. The modified portion detaches from the nucleus and acts as a cortical microtubule organizing center, while the rest of the spindle pole body remains at the nucleus.

Actins↗

Heterologous expression of mating-type genes in filamentous fungi.

Podospora anserina and Neurospora crassa, two filamentous heterothallic ascomycetes, have a single mating-type locus with two alternate forms called mat+ and mat- and A and a, respectively. Mating type controls entry into the sexual cycle, events subsequent to fertilization, and, in N. crassa, prevents the formation of mixed mating-type heterokaryons. The mating types of these two organisms display similarity in their DNA structure and in the encoded polypeptides involved in fertilization. Here we show that this molecular similarity reflects a functional homology with respect to mating identity. Transformation experiments show that the N. crassa mating-type genes can provide the fertilization functions in P. anserina strains devoid of mating specificity as well as in mat+ and mat- strains. Reciprocally, the introduction of P. anserina mating-type genes confers mating activity in N. crassa. Functional identity between the mating types is not observed for vegetative incompatibility or for post-fertilization events such as meiosis and ascosporogenesis.

DNA-Binding Proteins↗

Correlation between pairing initiation sites, recombination nodules and meiotic recombination in Sordaria macrospora.

The decrease of meiotic exchanges (crossing over and conversion) in two mutants of Sordaria macrospora correlated strongly with a reduction of chiasmata and of both types of "recombination nodules." Serial section reconstruction electron microscopy was used to compare the synapsis pattern of meiotic prophase I in wild type and mutants. First, synapsis occurred but the number of synaptonemal complex initiation sites was reduced in both mutants. Second, this reduction was accompanied by, or resulted in, modifications of the pattern of synapsis. Genetic and synaptonemal complex maps were compared in three regions along one chromosome arm divided into well marked intervals. Reciprocal exchange frequencies and number of recombination nodules correlated in wild type in the three analyzed intervals, but disparity was found between the location of recombination nodules and exchanges in the mutants. Despite the twofold exchange decrease, sections of the genome such as the short arm of chromosome 2 and telomere regions were sheltered from nodule decrease and from pairing modifications. This indicated a certain amount of diversity in the control of these features and suggested that exchange frequency was dependent not only on the amount of effective pairing but also on the localization of the pairing sites, as revealed by the synaptonemal complex progression in the mutants.

Ascomycota↗

Indirect intergenic suppression of a radiosensitive mutant of Sordaria macrospora defective in sister-chromatid cohesiveness.

Six ultra violet (UV) mutageneses were performed on the spo76 UV-sensitive mutant of Sordaria macrospora. Spo76 shows an early centromere cleavage associated with an arrest at the first meiotic division and therefore does not form ascospores. Moreover, it exhibits altered pairing structure (synaptonemal complex), revealing a defect in the sister-chromatid cohesiveness. From 37 revertants which partially restored sporulation, 34 extragenic suppressors of spo76 were isolated. All suppressors are altered in chromosomal pairing but, unlike spo76, show a wild type centromere cleavage. The 34 suppressors were assigned to six different genes and mapped. Only one of the suppressor genes is involved in repair functions.

Ascomycota↗

Defective pairing and synaptonemal complex formation in a Sordaria mutant (spo44) with a translocated segment of the nucleolar organizer.

The recessive meiotic mutant spo44 of Sordaria macrospora, with 90% ascospore abortion, exhibits striking effects on recombination (67% decrease), irregular segregation of the almost unpaired homologues, and a decrease in chiasma frequency in the few cases where bivalents are formed. Three-dimensional reconstructions of ten prophase nuclei indicate that pairing, as judged by the absence of fully formed synaptonemal complexes (SC), is not achieved although lateral elements (LE) assemble. The pairing failure is attributable to defects in the alignment of homologous chromosomes. The leptotene alignment seen in the wild type before SC formation was not observed in the spo44 nuclei. Dense material, considered to be precursor of SC central elements, was found scattered among the LE in two nuclei. The behaviour of spo44 substantiates the hypothesis that chromosome matching and SC formation are separable events. - The total length of the LE in the mutant is the same as in the wild type, but due to variable numbers and length of the individual LE, homologues cannot be lined up. Light microscopic observations indicate that the irregular length and number of LE is due to extensive chromosome breakage. The wild-type function corresponding to spo44 is required for both LE integrity and chromosome matching. Reconstructions of heterozygous nuclei reveal the presence of a supernumerary nucleolar organizer in one arm of chromosome 7. It is suggested that rDNA has been inserted into a gene whose function is involved in pairing or into a controlling sequence that interacts with the pairing process.

Ascomycota↗

The synaptonemal complex and the spindle plaque during meiosis in yeast.

Meiosis in Saccharomyces cerevisiae proceeds principally in the same manner as in other Ascomycetes. Leptotene is characterized by unpaired lateral components and pachytene by the presence of extensive synaptonemal complexes. The synaptonemal complex has the same dimensions and is similar in structure to those described for other organisms. Chromosome counts can now be made by reconstructing the synaptonemal complexes. Diplotene nuclei consistently contain a single polycomplex. The behaviour, doubling and the fine structure of the spindle plaque provide additional markers for the different stages of meiosis.

Cell Nucleolus↗