Sex-linked recessive lethal test of Drosophila melanogaster after exposure to 50-Hz magnetic fields.

To determine whether a 50-Hz magnetic field will induce mutations, a sex-linked recessive lethal test of Drosophila melanogaster was performed. Adult flies were exposed at an rms flux density of 500 mu T or 5 mT to the homogeneous field of a Helmholtz coil. The ambient field to which controls were exposed was less than 1 mu T. Exposures took place continuously for 13 to 14 days, which correspond to the life cycle of Drosophila at 25 degrees C. About 10,000 X-chromosomes were tested at each flux density. Recessive lethal mutation rates of 0.13, 0.21, and 0.18 percent were observed, respectively, for control, 500-mu T, and 5-mT conditions. By the Kastenbaum-Bowman significance test, the recessive lethal mutation rates in the 500-mu T and 5-mT conditions did not differ from the mutation rate of controls.     

The effect of population density on the elimination dynamics of a recessive lethal mutation from experimental populations

Dynamics of the elimination of the temperature-sensitive lethal mutation l(2)M167DTS from experimental populations of Drosophila melanogaster under permissive conditions (25 degrees C) was studied. We have shown a rapid elimination of the mutation from the populations, selection for fitness of heterozygous individuals, association of the selection with high larval density and with the direction of the cross that had produced the founder males of the l(2)M167DTS/+ population, and the effect of relative competitive ability of l(2)M167DTS/+ males on the efficiency of the mutation introduction. Modification systems were shown be involved in fitness selection under conditions of high larval density.     

A cell cycle G0-ts mutant, tsJT60, becomes lethal at the nonpermissive temperature after transformation with adenovirus 12 E1B 19K mutant

tsJT60, a temperature-sensitive (ts) cell-cycle mutant of Fischer rats, is viable at both the permissive (34 degrees C) and nonpermissive (40 degrees C) temperatures. The cells grow normally in exponential growth phase at both temperatures, but when stimulated with serum from G0 phase they enter S phase at 34 degrees C but not at 40 degrees C. tsJT60 cells transformed with human adenovirus (Ad) 12 dl205, which lacks the E1B 19-kDa polypeptide gene, were lethal at 40 degrees C, whereas tsJT60 cells transformed with Ad12 wt, dl207, which lacks E1B 58-kDa protein gene, or in206B, which produces 19- to 58- kDa fused protein, were viable. Degradation of cell DNA occurred in dl205-transformed tsJT60 cultured at both 34 degrees C and 40 degrees C. Neither cytocidal phenotype nor degradation of DNA occurred in 3Y1 cells (a parental line of tsJT60) transformed with dl205. These results suggest that the lethal phenotype and degradation of DNA are related to the ts mutation in tsJT60 and also to the lack of Ad12 E1B 19kDa polypeptide.     

A genetic suppressor of two dominant temperature-sensitive lethal proteasome mutants of Drosophila melanogaster is itself a mutated proteasome subunit gene

Two dominant temperature-sensitive (DTS) lethal mutants of Drosophila melanogaster are Pros26(1) and Prosbeta2(1), previously known as DTS5 and DTS7. Heterozygotes for either mutant die as pupae when raised at 29 degrees , but are normally viable and fertile at 25 degrees . Previous studies have identified these as missense mutations in the genes encoding the beta6 and beta2 subunits of the 20S proteasome, respectively. In an effort to isolate additional proteasome-related mutants a screen for dominant suppressors of Pros26(1) was carried out, resulting in the identification of Pros25(SuDTS) [originally called Su(DTS)], a missense mutation in the gene encoding the 20S proteasome alpha2 subunit. Pros25(SuDTS) acts in a dominant manner to rescue both Pros26(1) and Prosbeta2(1) from their DTS lethal phenotypes. Using an in vivo protein degradation assay it was shown that this suppression occurs by counteracting the dominant-negative effect of the DTS mutant on proteasome activity. Pros25(SuDTS) is a recessive polyphasic lethal at ambient temperatures. The effects of these mutants on larval neuroblast mitosis were also examined. While Prosbeta2(1) shows a modest increase in the number of defective mitotic figures, there were no defects seen with the other two mutants, other than slightly reduced mitotic indexes.     

tsJT60, a cell cycle G0-ts mutant, becomes lethal at non-permissive temperature by transformation with adenovirus 5 when the expression of E1B gene is lacking

tsJT60, a temperature-sensitive (ts) mutant cell line of Fischer rat, is viable at both permissive (34 degrees C) and non-permissive (39.5 degrees C) temperatures. The cells grow normally in exponential growth phase at both temperatures, but when stimulated with fetal bovine serum (FBS) from G0 phase they re-enter S phase at 34 degrees C but not at 39.5 degrees. When tsJT60 cells were transformed with adenovirus (Ad) 5 wild type, they grew well at both temperatures, expressed E1A and E1B genes, and formed colonies in soft agar. When tsJT60 cells were transformed with Ad5 dl313, that lacks E1B gene, the transformed cells grew well at 34 degrees C but failed to form colony in soft agar. They died very soon at 39.5 degrees C. 3Y1 cells (a parental line of tsJT60) transformed with dl313 grew well at both temperatures, although neither expressed E1B gene nor formed colonies in soft agar. The phenotype of being lethal at 39.5 degrees C of dl313-transformed tsJT60 cells was complemented by cell fusion with 3Y1BUr cells (5-BrdU-resistant 3Y1), but not with tsJT60TGr cells (6-thioguanine resistant tsJT60). These results indicate that the lethal phenotype is related to the ts mutation of tsJT60 cells and also to the deletion of E1B gene of Ad5.     

Adaptation to sublethal environmental stresses protects Listeria monocytogenes against lethal preservation factors.

A sublethal dose of ethanol (5%, vol/vol), acid (HCl, pH 4.5 to 5.0), H2O2 (500 ppm), or NaCl (7%, wt/vol) was added to a Listeria monocytogenes culture at the exponential phase, and the cells were allowed to grow for 1 h. Exponential-phase cells also were heat shocked at 45 degrees C for 1 h. The stress-adapted cells were then subjected to the following factors at the indicated lethal levels--NaCl (25%, wt/vol), ethanol (17.5%, vol/vol), hydrogen peroxide (0.1%, wt/vol), acid (pH 3.5), and starvation on 0.1 M phosphate buffer at pH 7.0 (up to 300 h). Viable counts of the pathogen, after the treatment, were determined on Trypticase soy agar-yeast extract, and survivor plots were constructed. The area (h.log10 CFU/ml) between the control and treatment curves was calculated to represent the protective effect resulting from adaptation to the sublethal stress factor. Adaptation to pH 4.5 to 5.0 or 5% ethanol significantly (P < 0.05) increased the resistance of L. monocytogenes to lethal doses of acid, ethanol, and H2O2. Adaptation to ethanol significantly (P < 0.05) increased the resistance to 25% NaCl. When L. monocytogenes was adapted to 500 ppm of H2O2, 7% NaCl, or heat, resistance of the pathogen to 1% hydrogen peroxide increased significantly (P < 0.05). Heat shock significantly (P < 0.05) increased the resistance to ethanol and NaCl. Therefore, the occurrence of stress protection after adaptation of L. monocytogenes to environmental stresses depends on the type of stress encountered and the lethal factor applied. This "stress hardening" should be considered when current food processing technologies are modified or new ones are developed.     

Acclimation effects on critical and lethal thermal limits of workers of the Argentine ant, Linepithema humile

For the Argentine ant Linepithema humile, bioclimatic models often predict narrower optimal temperature ranges than those suggested by behavioural and physiological studies. Although water balance characteristics of workers of this species have been thoroughly studied, gaps exist in current understanding of its thermal limits. We investigated critical thermal minima and maxima and upper and lower lethal limits following acclimation to four temperatures (15, 20, 25, 30 degrees C; 12L:12D photoperiod) in adult workers of the Argentine ant, L. humile, collected from Stellenbosch, South Africa. At an ecologically relevant rate of temperature change of 0.05 degrees Cmin(-1), CTMax varied between 38 and 40 degrees C, and CTMin varied between 0 and 0.8 degrees C. In both cases the response to acclimation was weak. A significant time by exposure temperature interaction was found for upper and lower lethal limits, with a more pronounced effect of acclimation at longer exposure durations. Upper lethal limits varied between 37 and 44 degrees C, whilst lower lethal limits varied between -4 and -10.5 degrees C, with an acclimation effect more pronounced for upper than lower lethal limits. A thermal envelope for workers of the Argentine ant is provided, demonstrating that upper thermal limits do likely contribute to distributional limits, but that lower lethal limits and limits to activity likely do not, or at least for workers who are not exposed simultaneously to the demands of load carriage and successful foraging behaviour.     

Recovery from potentially lethal damage of irradiated L5178Y-R and L5178Y-S cells: the effect of temperature and benzamide

Mouse lymphoma L5178 Y-S and Y-R cells differing in radiosensitivity by 1.5 times were treated with benzamide, an inhibitor of poly(ADP-ribosylation), for 24 h before and 18 h after X-irradiation, and incubated after irradiation at 25 degrees C and 37 degrees C. Clonogenic capacity of LY-S cells incubated at 25 degrees C exceeded that of the same cells incubated at 37 degrees C; the clonogenic capacity of LY-R cells did not vary with the postirradiation incubation temperature. Benzamide increased equally the radiosensitivity of LY-R cells incubated at both temperatures, whereas that of LY-S cells was only increased at 37 degrees C. Repair of potentially lethal damages to LY-S cells incubated at 25 degrees C was independent of the effectiveness of poly(ADP-ribosylation).     

Conditionally lethal and recessive UGA-suppressor mutations in the prfB gene encoding peptide chain release factor 2 of Escherichia coli.

Strains carrying mutations in the prfB gene encoding peptide chain release factor 2 of Escherichia coli were isolated. prfB1, prfB2, and prfB3 were selected as suppressor mutations of a lacZ (UGA) mutation at 37 degrees C, one of which, prfB2, is temperature sensitive in growth. A prfB286 strain was selected as a conditionally lethal mutant which grows at 32 but not at 43 degrees C and was shown to have UGA-suppressor activity. All the mutations are recessive UGA-suppressors. These data indicate that release factor 2 is essential to E. coli growth and that all mutants isolated here trigger suppression of the UGA codon.     

Gametic imprinting and expression of the gene Fused in the house mice

The discovery of the gametic imprinting phenomena in mammals makes it possible to have a new look at some facts concerning the expression and inheritance of genes with variable penetrance. Fused (Fu) is a dominant mutation located on chromosome 17, one of the few examples uses to demonstrate gametic imprinting in mice. This mutation has maternal effect connected with decrease in its penetrance. It was shown that t12 haplotype significantly reduces penetrance of the Fu in the progeny of Fu/t12 females. The results of reciprocal crosses of heterozygotes for t12 haplotypes indicate that penetrance of maternal Fu gene is reduced. Far more strong influence on the fused penetrance have the dominant suppressors, located beyond the chromosome 17. The penetrance of the fused gene decreases in that case up to 8-17%. Results of the experiments show strong influence of gametic pathway on penetrance of the gene by which the gene is transmitted to the next generation. The results also made it possible to describe the regularities of gametic imprinting. This phenomenon clearly indicates the existence of gametic and zygotic ontogenetic phases. According to the hypothesis proposed gametic phase of ontogenesis in mammals starts after initialization, i. e. after a process of chromosome erasing from epigenetic information and preparing to enter the next ontogenetic cycle.     

Frequency of mutations related to hereditary haemochromatosis in northwestern Poland

Hereditary haemochromatosis has been linked with C282Y and H63D mutations of the HFE gene. In Europe, frequencies of these mutations are the highest in Northern European countries and gradually decrease southwards. We analysed the prevalence of HFE mutations in 1517 DNA samples, including 1000 samples from the general population (subjects registered at general practitioner practices) in northwestern Poland, and 517 samples of cord blood from the same region. We identified 2 (0.13%) homozygotes and 117 (7.8%) heterozygotes for the C282Y mutation. As regards the H63D mutation (1505 DNA samples analysed), 38 (2.5%) samples were homozygotes and 380 (25%) were heterozygotes. Twenty-one (1.4%) compound heterozygotes were found. These results correspond well with data from other Central European countries and seem to confirm the hypothesis of North-South spread of the C282Y mutation.     

The Ash-1, Ash-2 and Trithorax Genes of Drosophila Melanogaster Are Functionally Related

Mutations in the ash-1 and ash-2 genes of Drosophila melanogaster cause a wide variety of homeotic transformations that are similar to the transformations caused by mutations in the trithorax gene. Based on this similar variety of transformations, it was hypothesized that these genes are members of a functionally related set. Three genetic tests were employed here to evaluate that hypothesis. The first test was to examine interactions of ash-1, ash-2 and trithorax mutations with each other. Double and triple heterozygotes of recessive lethal alleles express characteristic homeotic transformations. For example, double heterozygotes of a null allele of ash-1 and a deletion of trithorax have partial transformations of their first and third legs to second legs and of their halteres to wings. The penetrance of these transformations is reduced by a duplication of the bithorax complex. The second test was to examine interactions with a mutation in the female sterile (1) homeotic gene. The penetrance of the homeotic phenotype in progeny from mutant mothers is increased by heterozygosis for alleles of ash-1 or ash-2 as well as for trithorax alleles. The third test was to examine the interaction with a mutation of the Polycomb gene. The extra sex combs phenotype caused by heterozygosis for a deletion of Polycomb is suppressed by heterozygosis for ash-1, ash-2 or trithorax alleles. The fact that mutations in each of the three genes gave rise to similar results in all three tests represents substantial evidence that ash-1, ash-2 and trithorax are members of a functionally related set of genes.     

Temperature-Sensitive Mutations in DROSOPHILA MELANOGASTER. IX. Dominant Cold-Sensitive Lethals on the Autosomes

Ethyl methanesulfonate-treated autosomes were screened for the presence of dominant cold-sensitive (DCS) lethal mutations in Drosophila melanogaster. None was found among 6,552 treated and 168 untreated third chromosomes. Twenty-three DCS-L chromosomes which caused death at 17 degrees C but survived at 22 degrees C and 29 degrees C were recovered from 5,046 mutagenized chromosome 2's.-The DCS-L mutations all mapped around dp and appeared to be functionally allelic. Lethality of heterozygotes for most of the DCS-L's occurred over a prolonged interval from the embryonic through the larval instars. Prolonged incubation at 17 degrees C did not demonstrate any maternal effect on zygotic survival.     

Fertility of Drosophila melanogaster females affected by mutation l(2)M167DTS

We studied the fertility of D. melanogaster females heterozygous for the dominant temperature sensitive mutation l(2)M167DTS, which exerts a recessive lethal effect at 25 degrees C, under the conditions of stable temperature regimes 25, 28, and 29 degrees C and changing regimes 25-->29 degrees C and 29-->25 degrees C. It was shown that inhibition of total activity of oogenesis due to a decreased number of functioning ovarioles is one of the mechanisms underlying the decreased fertility of l(2)M167DTS /+ females. Analysis of individual fertility of each female confirmed also the role of sterility as a component of fertility of the females. Sterilization was realized due both to full depletion of functioning ovarioles and disturbed mechanism of laying the mature eggs onto a substrate as a result of violation of the feedback blocking normal ovulation, which led to the breakdown of ovarioles and filling of the abdominal cavity with mature oocytes. A significant polymorphism of heterozygous females by their fertility was observed. The intensity of sterilization and mortality of l(2)M167DTS/+ females sharply increased at an elevated temperature (29 degrees C), especially at the pupal stage.     

A transposon insertion in the Arabidopsis SSR16 gene causes an embryo-defective lethal mutation

The SSR16 gene of Arabidopsis has been identified as a gene encoding a ribosomal protein S16 homolog through analysis of a transposon insertion mutation. The insertion mutation is lethal, arresting embryonic development at approximately the transition from the globular to the heart stage of embryonic development. Co-segregation of the mutant phenotype with the transposon-borne drug-resistance marker and loss of the inserted transposon concomitant with phenotypic reversion provided evidence that the transposon had caused the mutation. Sequences flanking the insertion site were amplified from DNA of viable heterozygotes by thermal asymmetric interlaced (TAIL) PCR. The amplified fragment flanking the 3' end of the inserted element was sequenced and found to be identical to an Arabidopsis expressed sequence tag (EST). The EST, in turn, contained a coding sequence homologous to the ribosomal protein S16 (RPS16) of bacteria such as Escherichia coli, Bacillus subtilis and Salmonella typhimurium , as well as Neurospora crassa mitochondria and higher plant plastids. Thus the gene identified by the embryo-defective lethal insertion mutation encodes an RPS16 homolog and has been designated the SSR16 gene.     

A conditionally lethal mutation of Escherichia coli affecting the gene coding for ribosomal protein S2 (rpsB).

Localized P1 mutagenesis has been used to isolate conditionally lethal mutations in the four-minute region of the Escherichia coli genome. One such mutation, ts25, has been mapped at about 3.7 minutes between the popC and dapD genes. This mutation leads to thermosensitivity of growth and impaired in vivo assembly of 30 S ribosomal subunits at 42 °C. The strain carrying the mutation has an altered S2 ribosomal protein as judged by (1) its inability to maintain stable complex with the ribosome under mild washing conditions and (2) its altered electrophoretic mobility.Spontaneous reversion to temperature independence can restore both the normal assembly in vivo of 30 S ribosomal subunits at 42 °C and the normal electrophoretic behaviour of the S2 ribosomal protein in vitro.We conclude therefore that the ts25 mutation affects the structural gene for ribosomal protein S2 (rpsB).     

Heterosis in a wild strain of Drosophila polymorpha with a lethal closely linked to the major esterase locus

A laboratory population derived from a single wild inseminated female has a lethal so closely linked to the major esterase locus that in 3 years of observations no crossover products have been detected. Linkage with a chromosome inversion was excluded by cytological analysis. The heterozygotes are superior to the homozygotes in egg-adult viability, egg-laying rate, and longevity. Electrophoretic analysis of larvae, pupae, and 2-h-old adults shows that the specific phase for the lethal effect in homozygotes is the pupal stage.     

Differentiation of the effects of lethal pH and water activity: food safety implications

The influence of a second lethal stress (SLS) was investigated when populations of Escherichia coli M23 OR.H- were exposed to either a low water activity (aw) of 0.90 or a pH of 3.50 after 24 h at 25 degrees C. Regardless of the initial stress, E. coli M23 OR.H- populations initially demonstrated biphasic inactivation kinetics consisting of a rapid first phase of death followed by a slower second phase. When cultures initially exposed to aw 0.90 experienced an SLS of pH 3.50, a second rapid inactivation period was observed before a subpopulation of more resistant cells emerged. This subpopulation was able to persist for approximately 50 h after imposition of the SLS. In contrast, E. coli M23 OR.H- cells first exposed to a pH of 3.50 were inactivated rapidly to levels below the limits of detection upon imposition of an SLS of aw 0.90. It is hypothesized that pH stress constitutes a large energy drain on the cell and subsequently sensitizes it to other environmental constraints requiring expenditure of metabolic energy.     

Dominant lethal mutations in the dnaB helicase gene of Salmonella typhimurium

A class of dominant lethal mutations in the dnaB (replicative helicase) gene of Salmonella typhimurium is described. The mutated genes, when present on multicopy plasmids, interfered with colony formation by Escherichia coli host strains with a functional chromosomal dnaB gene. The lethal phenotype was expressed specifically in supE (glutamine-inserting) host strains and not in Sup+ strains, because the mutant genes, by design, also possessed an amber mutation derived from a glutamine codon. Mutations located at 11 sites by deletion mapping and DNA sequence analysis varied in the temperature dependence and severity of their lethal effects. None of the mutations complemented a dnaB(Ts) host strain at high temperature (42 degrees C). Therefore, these nonfunctional DnaB proteins must engage some component(s) of the DNA replication machinery and inhibit replication. These mutations are predicted to confer limited, specific defects in either the catalytic activity of DnaB or the ability of DnaB to interact with one of its ligands such as DNA, nucleotide, or another replication protein. The variety of mutant sites and detailed phenotypes represented in this group of mutations may indicate the operation of more than one specific mechanism of lethality.     

Expression of a new mutation (Axd) causing axial defects in mice correlates with maternal phenotype and age

A new autosomal mutation, Axd (axial defects), is described. Axd segregates in a simple Mendelian fashion, and it is dominant with incomplete penetrance and variable expressivity. The phenotype of Axd heterozygotes ranges from a variety of tail anomalies to visibly normal tails. Approximately 12% of neonates from curly-tail (CT) F1 (Axd/+) x F1 (Axd/+) matings exhibit open neural tube defects (NTD) in the lumbosacral region and 16% have curly tails. Mean litter sizes and resorption rates comparable to wild type indicate that homozygosity for Axd is not obligately lethal. Genetic background plays a major role in Axd expression. Strains such as BALB/cByJ allow the highest penetrance of the mutation in single dose (46%), whereas, in CF-1 mice Axd is recessive. The tail phenotype of heterozygous Axd/+ dams, in part reflective of their genetic background, correlates with the incidence of NTD in F2 offspring: CT mothers produce significantly more neonates with frank NTD than normal tail mothers. At the one embryonic period examined for this study (D13/D14 post-coitus), an 85% higher incidence of total axial defects is observed than among the F2 at birth. Unchanging litter size and the relative increase in phenotypically normal offspring by birth suggest that Axd acts by delaying posterior neural tube closure. One of the most significant findings in this study is that maternal age influences the survival of Axd embryos in utero. Axd/+ dams older than 8 months yield fewer mean implants, higher resorption rates, and fewer viable embryos with axial defects than do Axd/+ dams younger than 8 months. Axd is not allelic to nor linked to the Sp (splotch) gene which also affects neurulation.