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PubMed1980

Disproportionate rDNA replication does occur in diploid tissue in Drosophila hydei.

The rDNA content in Drosophila hydei has been compared in wild-type and in two translocation genotypes possessing only one nucleolus organizer. In highly polyploid salivary glands where rDNA is underreplicated, an 'independent polytenization' of the rDNA occurs resulting in about the same rDNA level in each genotype independently of the number of nucleolous organizers present in the genome. Thus, the situation in the salivary glands of D. hydei is similar to that in D. melanogaster (Spear and Gall 1973). In tetraploid thoracic muscle where rDNA is not underreplicated, the rDNA percentage in the two translocation genotypes is also considerably increased, although the wild-type level is not completely attained. This result shows that rDNA replication is independently controlled even in a non-underreplicating tissue. In larval diploid brain the situation in the two translocation stocks is dissimilar: in one genotype the rDNA content remains unaltered whereas in the other it is increased. This demonstrates for the first time that a gene compensation does occur in a diploid tissue.

Animals↗

PubMed1975

Strains of Escherichia coli diploid for the chromosomal origin of DNA replication.

F' strains of E. coli have been isolated which are merodiploid for various chromosomal segments between 66 and 78 minutes. Strains diploid for the chromosomal DNA between the genes bgl and mtl grow slowly, have a reduced DNA/mass and an increased cell size. These properties could result if the chromosomal replication origin and a second, extrachromosomal, copy of the origin (located in this case on the F') were to compete for a substance required to initiate replication. We therefore suggest that these strains are diploid for the chromosomal origin of replication and that, therefore, the origin is located between bgl and mtl, that is between 71 and 73 minutes on the E. coli chromosome.

Bacterial Proteins↗

PubMed1983

Interferon induction of (2'-5') oligoisoadenylate synthetase in diploid and trisomy 21 human fibroblasts: relation to dosage of the interferon receptor gene (IRFC).

Trisomy 21 human fibroblasts are more sensitive to human interferon-alpha (IFN-alpha) than are diploid controls, consistent with the location of the gene (IFRC) which codes for the IFN-alpha receptor on chromosome 21. When compared in the antiviral assay, the difference in sensitivity is five- to tenfold, much greater than the 50% difference in IFRC gene dosage. An understanding of the mechanism by which this amplification of gene dosage occurs is relevant to the specific pathology of Down's syndrome and as a model system for studying the pathogenic effects of chromosomal aneuploidy. The enzyme (2'-5') oligoisoadenylate synthetase (2-5A synthetase), which is believed to be central to the interferon-induced antiviral response, is induced 50% more in trisomy 21 fibroblasts than in diploid controls. Thus the amplification in response occurs subsequent to the binding of IFN-alpha to its receptor and the triggering of the first set of intracellular events, the latter exemplified by the induction of 2-5A synthetase. Similar results were obtained with IFN-gamma, consistent with other evidence which indicates that a gene coding for a separate IFN-gamma receptor is also located on chromosome 21.

Diploidy↗

PubMed1976

Unintegrated ribosomal genes in diploid and polytene tissues of Drosophila melanogaster.

We have examined DNA from polytene salivary glands and diploid brains and imaginal discs of male and female larvae having one or two nucleolus organizers. DNA having an estimated molecular weight of 5 X 10(9) or greater was obtained by sucrose gradient sedimentation of gently prepared lysates. Hybridization of the gradient fractions with 3H-ribosomal RNA REVEALS THAT 42% OF THE RIBOSOMAL GENES ARE FOUND IN DNA of lower molecular weight (approximately 3 X 10(8) daltons) in the salivary glands of every genotype examined. In the brains and imaginal discs, by contrast, all of the ribosomal genes are found in the high molecular weight peak except in females with one nucleolus organizer where 42% are found in lower molecular weight DNA, as in the salivary gland. Thus unintegrated genes are not an exclusive feature of polytene tissue, but can occur in diploid tissue as well in at least one genotype.

Animals↗

PubMed1979

Non-repetitive DNA sequence divergence in phylogenetically diploid and tetraploid teleostean species of the family cyprinidae and the order isospondyli.

Non-repetitive DNA of anciently tetraploid teleostean species was analysed for the presence of duplicated sequences. Closely related diploid species were investigated in comparison. From the reassociation kinetics of total nuclear DNA, rate constants and fraction sizes of classes of repetitive and non-repetitive sequences were determined. DNA fractions enriched in the slowest renaturing sequence class were determined. DNA fractions enriched in the slowest renaturing sequence class were prepared and subjected to reassociation. The rate constants of these reactions were compared with the values expected for single-copy DNA from analytical genome size determinations. From reassociated DNA enriched in non-repetitive sequences also the melting temperatures were determined as a measure of internal base sequence heterogeneity. It has been shown that the two ancient tetraploids Cyprinus carpio and Thymallus thymallus are, with regard to the thermal stability of reassociated non-repetitive DNA, and with regard to the correspondence of reaction rates with the values expected for single copy DNA, indistinguishable from diploid controls (Rutilus rutilus, Clupea harengus and Sprattus sprattus). The tetraploid species Salmo irideus, Salvelinus fontinalis and Coregonus lavaretus appear as very recent tetraploids with regard to these criteria. The significance of the results for estimating the time of occurrence of polyploidisation events in these taxa is discussed.

Animals↗

PubMed1976

Haplo-diploid locust embryos arising by accidental thelytoky in Chortoicetes terminifera investigated by G-banding.

Two mosaic sibling embryos of the Australian plague locust, Chortoicetes terminifera are reported with haploid and diploid cell lines in widely differing proportions. One small chromosome pair involved in the two cases has alternative morphology and a B-chromosome is present in one. In addition, G-banding identifies two medium-sized chromosome pairs and alternative states of a second small pair. Using these markers it is clear that both diploid cell lines are homozygous for the chromosomes of the corresponding haploid line. These embryos have thus developed by accidental parthenogenesis from haploid cells, some of which were duplicated by endomitosis after development began.

Animals↗

PubMed1991

Synchronized meiosis and recombination in fission yeast: observations with pat1-114 diploid cells.

The mutation pat1-114 has been used to synchronize meiosis in the fission yeast Schizosaccharomyces pombe. We have investigated several aspects of such synchronized meiotic cultures. In both pat1-114 and pat1+ diploids, meiotic landmark events are initiated at the same time after meiosis induction, but synchrony is much more pronounced in the pat1-114-driven meiosis. Commitment to recombination and to meiosis have been timed at 2 h after meiotic induction. Due to a seven-fold reduction of intragenic recombination frequency in the ade6 region of pat1-114 diploids, physical analysis of recombination has not been possible. We have distinguished three factors that influence intragenic recombination frequencies: temperature, azygotic versus zygotic meiosis, and the nature of the pat1 allele. Differences and similarities in the timing of meiotic landmarks in S. cerevisiae and S. pombe are discussed.

Cell Cycle↗

PubMed1990

p-fluoro-phenylalanine resistance in Aspergillus nidulans diploid cells: evidence that dominant, lethal mutations are involved.

An unexpectedly large number of p-fluoro-phenylalanine (FPA)-resistant mutants have been recovered after UV-irradiation of wild type diploid conidia of Aspergillus nidulans. At least five different classes of mutants, possibly corresponding to five different loci, have been identified. Two of them may be the dominant loci which have already been described but the others (a minimum of three loci) are completely different. Mutations in these loci confer high level FPA resistance in the heterozygous diploids, being lethal in the haploids; one mutation has been preliminarily mapped to chromosome I and another to chromosome III.

Aspergillus nidulans↗

PubMed1982

Direct selection of complementing diploids from PEG-induced fusion of Bacillus subtilis protoplasts.

A minimal medium containing horse serum is described on which Bacillus subtilis protoplasts revert to bacillary forms at high frequency (ca. 30%). Used as a plating medium for a mixture of polyethyleneglycol-treated protoplasts from two complementary polyauxotrophic parental strains, it selects the prototrophic fusion products efficiently, and also allows isolation of various auxotrophic recombinants. These prototrophs and recombinants amount respectively to 1% and 10% of the regenerated bacteria. We confirm that two types of prototrophs can be isolated after fusion: stable recombinants and complementing diploids, the latter segregating into various types of recombinants. Based on easily recognized colonial aspects, an approximate estimation of the proportion of the two types becomes possible when a spoOA mutation has been introduced in one of the parents. At least 50% of the prototrophic fusion products are complementing diploids. Incidently, the data also settle a controversy by showing the dominance of spoOA mutations in heterozygotic bacteria.

Bacillus subtilis↗

PubMed1988

Further studies on recombination in diploid clones from Bacillus subtilis protoplast fusion.

Diploid prototrophs were obtained from protoplast fusion of Bacillus subtilis strains. They are unstable but upon further cultivation they stabilize retaining diploidy but are genetically inactive. It has been suggested that recombination between the parental chromosomes is involved in the production of stable prototrophs and recombinants. In this work the occurrence of this recombination was searched for by determining genetic linkages in transformation experiments. In prototrophs two alleles: hisH2 and trpE8 carried originally on each parental chromosome, were shown to be 48% co-transformable in a stable clone whereas they were only cotransformed in 10% of the unstable colonies. For Trp- recombinants (the most frequent type of a Leu- Met- Thr- x Ade- Ura- Trp- fusion pair) lysed protoplasts were used as donor DNA for the transformations. High values of co-transfer for Ura+ Met+ were obtained. These results confirm the occurrence of recombination in stable diploid clones, prototrophs or recombinants.

Alleles↗

PubMed1975

Mutations affecting mitotic recombination frequency in haploids and diploids of the filamentous fungus Aspergillus nidulans.

A haploid strain of Asp. nidulans with a chromosome segment in duplicate (one in normal position on chromosome I, one translocated to chromosome II) shows mitotic recombination, mostly by conversion, in adE in a frequency slightly higher than in the equivalent diploid. A method has been devised, using this duplication, for the selection of rec and uvs mutations. Six rec mutations have been found which decrease recombination frequency in the haploid. One mutation selected as UV sensitive showed a hundred fold increase in recombination frequency in the haploid (pop mutation) and probably the same in diploids. The increased frequency is both in gene conversion and in crossing over, and the exchanges appear in clusters of two or more. pop is allelic to uvsB (Jansen, 1970) which had been found to affect mitotic but not meiotic recombination. It is suggested that mutations of this type interfere with the control mechanism which determines that high recombination is confirmed to the meiotic nuclei and avoided in somatic nuclei.

Aspergillus nidulans↗

PubMed1993

Self-diploidization in Saccharomyces cerevisiae kar2 heterokaryons.

Zygotes isolated by micromanipulation from crosses of Saccharomyces cerevisiae strains, one of which carries a kar mutation, give rise most frequently to cytoductant colonies showing the nuclear constitution of either one of the two haploid parental strains. In crosses of kar2-1 strains to wild-type, about 10% of the cytoductants of both mating types are homozygous autodiploids. There is evidence indicating that self-diploidization occurs by fusion between sibling nuclei in the heterokaryotic zygote. Here we describe this phenomenon and propose to take advantage of it for the construction of genotypically-defined diploids able to mate, and of polyploid strains, which are useful tools in genetic and cytological studies.

Crosses, Genetic↗

PubMed1996

DNA representation of variegating heterochromatic P-element inserts in diploid and polytene tissues of Drosophila melanogaster.

Position-effect variegation (PEV) is the mosaic expression of a euchromatic gene brought into juxtaposition with heterochromatin. Fourteen different transformed Drosophila melanogaster lines with variegating P-element inserts were used to examine the DNA levels of these transgenes. Insert sites include pericentric, telomeric and fourth chromosome regions. Southern blot analyses showed that the heterochromatic hsp26 transgenes are underrepresented 1.3- to 33-fold in polytene tissue relative to the endogenous euchromatic hsp26 gene. In contrast, the heterochromatic hsp26 transgenes are present in approximately the same copy number as the endogenous euchromatic hsp26 gene in diploid tissue. It appears unlikely that DNA loss could account for the lack of gene expression in diploid tissues seen with these examples of PEV.

ATP-Binding Cassette Transporters↗

PubMed1979

Alcohol dehydrogenase activity in diploid and autotetraploid Phlox.

The effect of polyploidy on the activity of alcohol dehydrogenase was examined in a series of diploid and synthetic autotetraploid Phlox drummondii. In most cases autotetraploids had about twice as much activity as the corresponding diploids, in two cases autotetraploids had about 1.5 times more activity, and in one wild seed pair the activity of the tetraploid was somewhat lower.

Alcohol Oxidoreductases↗

PubMed1976

Erythrocyte enzyme activities in diploid and triploid salamanders (Pleurodeles waltlii) of both sexes.

Studies of several enzyme activities and of some metabolites in erythrocytes of the newt Pleurodels waltilii are described. The results indicate that differences exist between enzyme activities in males and females and in diploid and triploid animals. Females have higher erythrocyte enzyme activities than males. However, triploid animals have lower enzyme activites in their erythrocytes than diploid animals; this observation is more striking in females than in males.

Alleles↗

PubMed1996

Comparison of the primary structure of waxy proteins (granule-bound starch synthase) between polyploid wheats and related diploid species.

The waxy proteins encoded by the genomes A, B, and D in polyploid wheats and related diploid species were isolated by SDS-PAGE. The N-terminal amino acid sequences of mature proteins and V8 protease-induced fragments were determined. A total of five amino acid substitutions was detected in these sequences, which represent about 10% of the whole sequences of the waxy proteins. A comparison of these sequences in polyploid wheats with those in related diploid species revealed the following: (i) waxy proteins encoded by the A genome of polyploid wheats were identical to that of Triticum monococcum, (ii) the waxy protein encoded by the B genome of T. turgidum was identical to that of T. searsii, but differed from those of T. speltoides and T. longissimum by one amino acid substitution, (iii) the waxy protein encoded by the B genome of T. aestivum differed from that encoded by the B genome of T. turgidum by one amino acid substitution, and (iv) the waxy protein encoded by the D genome of T. aestivum was identical to that of T. tauschii.

Amino Acid Sequence↗

PubMed1993

BCNU-sensitivity in parental cells and clones from four freshly resected near-diploid human gliomas: an astrocytoma, an anaplastic astrocytoma and two glioblastomas multiforme.

We compared the BCNU sensitivity of 4 freshly resected tumors (astrocytoma WM, and malignant gliomas MK, MB, and AM) and their clones to their karyology. The majority of primary cells in all 4 tumors had near-diploid chromosome numbers (2n +/-) and all were resistant to concentrations of BCNU exceeding 10 micrograms/ml. Following in vitro cultivation, the cells from tumors WM and MB retained their 2n +/- modal chromosome number with little change in the complexity of the karyotype. In contrast, tumors MK and AM demonstrated a more unstable genome. The modal chromosome number of MK shifted from 45 to 86 and that of tumor AM from 45 to 90. Karyotyping demonstrated additional ploidy changes and new marker chromosomes in both tumors. The colony forming assay (CFA) performed on the in vitro cultivated cells demonstrated little change in the sensitivity to BCNU in tumors WM and MB, while tumors MK and AM exhibited greater than a one log cell kill at 10.0 micrograms/ml and 15.0 micrograms/ml BCNU, respectively. The modal chromosome number and BCNU sensitivity followed a similar pattern in the 30 clones that were isolated; 21 clones with near-diploid and pseudodiploid chromosome numbers were all resistant to BCNU doses at or greater than 10 micrograms/ml. In contrast, 9 clones isolated from the 3 malignant gliomas with 3n +/- and 4n +/- modal chromosome numbers were sensitive to this concentration of BCNU. The karyotypes of the hyperdiploid clones were more complex; they contained 5 or more ploidy changes and/or had marker chromosomes. These studies confirm the association of diploidy and BCNU-resistance in freshly resected malignant gliomas.

Astrocytoma↗

PubMed1993

Overexpression of asparagine synthetase in albizziin-resistant murine diploid embryonic stem cells.

Gene amplification is commonly observed in primary tumors and established drug-resistant cell lines, both of which are generally aneuploid. However, this process is undetectable (frequency < 10(-9) in normal diploid mammalian cell lines. To investigate whether gene amplification can occur in pluripotent diploid cells, we have selected drug-resistant mutants of mouse embryonic stem (ES) cells. We had previously found that Chinese hamster ovary (CHO) and human cell lines selected in albizziin (Alb), an amino acid analog of L-glutamine, overexpress asparagine synthetase (AS) due to gene amplification. The same drug selection system was applied to ES cells to isolate single-step and multistep drug-resistant mutants. Albizziin-resistant ES cells exhibited elevated levels of AS; however, drug resistance in ES cells was associated with mRNA overexpression without gene amplification. AS gene amplification was observed in only one drug-resistant cell line and was preceded by AS mRNA overexpression. Gene amplification in the latter coincided with the loss of the pluripotent nature of the ES cells.

Amino Acids, Diamino↗