Determination of the probable ontogenic time of action of lethal factors on thesecond chromosome of Drosophila melanogaster derived from a natural population

From the study of the genetic load of second chromosome factors in a natural population of Drosophila melanogaster, 15 lethal-bearing strains were recovered and maintained in the laboratory balanced against Ins (2L+2R), Cy, L⁴. For each lethal factor, the probable time of action during development was determined by the appearance of a sharp reduction, at any given stage, in the frequency of individuals compared to that expected in the absence of the lethal factor. Carried out in this way, the analysis suggested that sevne were embryonic lethals, two larval lethals and three pupal lethals. Additionally, three gave no evidence of affecting any of the above-mentioned stages; these are interpreted as gametic lethals.     

Unstable ascospore color mutants of Ascobolus immersus

Summary In Ascobolus immersus, 16 unstable mutants with white ascospores were isolated. These mutants mutate back to the wild-type phenotype with brown ascospores. Only two mutants B and 301 are the object of the present study. At least in these two mutants it was demonstrated that true back-mutations occur. The frequencies of back-mutations are quite high and can reach 0.30 in the case of mutant B.The reversions appear at well defined but different for each mutant stages of the developmental cycle. These stages are relatively short and involve not more than about ten nuclear divisions cycles. In the case of mutant B, the back-mutations occur only in very young mycelia and at elevated temperature. Thus, the frequencies of back-mutations observed depend on the time of the transfer to higher temperature condition.The back-mutations of 301 mutant occur only in spermatized female organ and depend on the genotype of the female parent.The data established for Ascobolus immersus are compared with genic instability due to transposable elements described in corn by B. Mc Clintock.     

Cytogenetic Analysis of Chromosome 3 in DROSOPHILA MELANOGASTER: Mapping of the Proximal Portion of the Right Arm

In order to define more precisely the most proximal portion of chromosome 3R in Drosophila melanogaster, several new chromosome aberrations involving this region have been recovered and analyzed. These new arrangements were recovered as induced reversions of two dominant mutations, Antp^Ns and dsx^D, located in the region of interest. The results of the analysis have allowed the localization of several existing mutations, have further elucidated the complex homoeotic locus which resides in this region, and have confirmed the efficacy of this type of screen in the analysis of specific chromosome regions.     

Hypomorphic lethal mutations and their implications for the interpretation of lethal complementation studies in Drosophila

In a small region of the X chromosome of Drosophila melanogaster, we have found that a third of the mutations that appear to act as lethals in segmental haploids are viable in homozygous mutant individuals. These viable mutations fall into four complementation groups. The most reasonable explanation of these mutations is that they are a subset of functionally hypomorphic alleles of essential genes: hypomorphic mutations with activity levels above a threshold required for survival, but below twice that level, should behave in this manner. We refer to these mutations as "haplo-specific lethal mutations." In studies of autosomal lethals, haplo-specific lethal mutations can be included in lethal complementation tests without being identified as such. Accidental inclusion of disguised haplo-specific lethals in autosomal complementation tests will generate spurious examples of interallelic complementation.     

The cytogenetics of mutator gene-induced X-linked lethals in Drosophila melanogaster

A third chromosome mutator gene effectively increases the spontaneous frequency of sex-linked recessive lethals in females but not in females of Drosophila melanogaster. Approximately half the mutator-induced mutants occur as clusters of the same mutant implying a premeiotic origin. An appreciable number of the mutator-induced lethals are associated with comparatively long deficiencies of several salivary gland chromosome bands. The possible modes of mutator gene action are conjectured.     

Genetic Damage at Specific Gene Loci in Drosophila with Betatron X-Rays

The mutation rate was determined for mature sperm at eight specific gene loci on the third chromosome of Drosophila melanogaster using the low ion density radiations of 22 Mev betatron X-rays. A dose of 3000 rads of betatron X-rays produced a mutation rate of 4.36 x 10^-8 per rad/locus. Among the mutations observed, 66% were recessive lethals and 34% viable when homozygous. Only one of the 24 viable mutations was associated with a chromosome aberration. Among the 47 recessive lethals, no two-break aberrations were detected in 48.9% of the lethals, deletions were associated with 42.2%, inversions with 6.7% and translocations with 2.2%.—When these genetic results are compared to those for 250 KV X-rays, the mutation rate for betatron treatments was slightly lower (.76), the recessive lethal rate among induced mutations was higher, and the chromosome aberrations among lethal mutations were slightly lower than with 250 KV X-rays. Although the two types of irradiations differ by an ion density of approximately ten, the amount and types of inheritable genetic damage induced by the two radiations in mature sperm were not significantly different.     

The effect of caffeine on repair systems in oocytes of Drosophila melanogaster. I

Young Drosophila females were treated with caffeine, then mated for 24 h to males that had been irradiated with 2000 R X-irradiation, so that only mature spermatozoa were sampled. The radiation-induced frequency of dominant lethals and sex chromosome loss in the paternal genome was determined. The results show that treatment of females with caffeine leads to an increase in the frequencies of radiation-induced dominant lethals and to sex-chromosome loss.When young virgin females of the radioresistant stock RöI₂ were treated with caffeine and then irradiated with 3000 R X-irradiation, a striking increasein dominant lethals (in the maternal genome) was observed; caffeine treatment increased the X-ray response of the radioresistant stock to the level of the normal (+60) stock. It is suggested that caffeine reduces the efficiency of a system in Drosophila oocytes that repairs X-ray-produced chromosome breaks in both the paternal and maternal genomes.     

Temperature-Sensitive Mutations in DROSOPHILA MELANOGASTER. Xvii. Heat- and Cold-Sensitive Lethals on Chromosome 3

Ethyl methanesulfonate-treated third chromosome of Drosophila melanogaster were tested for the presence of dominant and recessive temperature-sensitive lethal mutations at 17 degrees , 22 degrees and 29 degrees C. Out of 1,176 chromosomes tested, no dominant ts lethals, 21 heat-sensitive, 22 cold-sensitive and 10 heat-cold-sensitive lethals were recovered. Heat-cold sensitivity was produced by a single mutation in all cases. Sixty-two percent of the ts lethals were fertile as homozygotes in both sexes. Surprisingly, 88% of the ts lethals mapped between st and Sb, a region straddling the centromere and estimated to comprise 12.9% of the genetic length and 55% of the physical length of chromosome 3. All but one of the heat- and cold-sensitive lethals complemented with each other at their respective restrictive temperatures.     

Genetics of Acetylcholinesterase in DROSOPHILA MELANOGASTER

Genes in Drosophila melanogaster that control acetylcholinesterase (AChE) were searched for by segmental aneuploidy techniques. Homogenates of flies containing duplications or deletions for different segments were assayed for enzyme activity. A region on the third chromosome was found for which flies having one does consistently gave lower AChE activity than euploid flies, which were in turn had lower activity than flies with three doses. The activity differences were in the approximate ratio 1:2:3. Fine structure deletion mapping within this region revealed a very small segment for which one-dose flies have approximately half-normal activity. To obtain putative AchE-null mutations, lethal mutations within this region were assayed. Four allelic lethals have approximately half-normal activity in heterozygous condition. These lethals probably define the structural locus (symbol: Ace) for AChE.     

The Frequency and Allelism of Lethal Chromosomes in Isolated Desert Populations of DROSOPHILA PSEUDOOBSCURA

Second-chromosome lethals were extracted from four populations of Drosophila pseudoobscura in Southern California. Two of the populations were from desert oases and two from the classic habitat on Mt. San Jacinto, previously studied by Dobzhansky. Allelism tests were made on the lethals within and between all locations. The frequency of lethal second-chromosomes in each location was 0.18, and this was not different from the results of other workers for samples throughout the species range. Interpopulational allelism rates were about 0.005, and not different from earlier results of Dobzhansky. Intrapopulational rates in this study were, with one exception, the same as the interpopulational rates, and significantly lower than Dobzhansky found using the third chromosome. This may be due to lethals being linked with heterotic third-chromosome inversions. The allelism rate of the exceptional population (about 0.03 and equal to Dobzhansky''s intrapopulational results) may be due to heterotic lethals, or a founder effect. Two lethals were found in three populations each, possibly due to migration among these populations, which are up to 334 km apart.     

Studies on the Sex-Specific Lethals of DROSOPHILA MELANOGASTER. V. Sex Transformation Caused by Interactions between a Female-Specific Lethal, Sxlf#1, and the Male-Specific Lethals mle(3)132, msl-227, and mle

Interactions between a female-specific lethal mutant, Sxl^f#1, and each of three male-specific lethal mutants, mle(3)132, msl-2²⁷ and mle, of Drosophila melanogaster were observed to produce morphological changes in various sexually dimorphic external characters. Chromosomal females heterozygous for Sxl^f#1 and homozygous for any one of the male-specific lethals (and to a lesser degree heterozygous for male-specific lethals) sometimes had sex combs, male-type tergites, male-type sternites, male-type anal plates or male-type external genitalia. Penetrance was not high and expression was often incomplete; single individuals never had all the sexually dimorphic structures transformed. When mothers were homozygous for male-specific lethals, higher proportions of female progeny were affected than when mothers were heterozygous, suggesting a maternal effect.     

Genetic Analysis of the Centromeric Heterochromatin of Chromosome 2 of DROSOPHILA MELANOGASTER: Deficiency Mapping of Ems-Induced Lethal Complementation Groups

Until recently, little was known of the genetic constitution of the heterochromatic segments of the major autosomes of Drosophila melanogaster . Our previous report described the genetic dissection of the proximal, heterochromatic region of chromosome 2 of Drosophila melanogaster by means of a series of overlapping deficiencies generated by the detachment of compound second autosomes (Hilliker and Holm 1975). Analysis of these deficiencies by inter se complementation, pseudo-dominance tests with proximal mutations and allelism tests with known deficiencies provided evidence for the existence of at least two loci between the centromere and the light locus in 2L and one locus in 2R between the rolled locus and the centromere. These data in conjunction with cytological observations demonstrated that light and rolled and three loci lying between them are located within the proximal heterochromatin of the second chromosome.——The present report describes the further analysis of this region through the induction with ethyl methanesulphonate (EMS) of recessive lethals allelic to the 2L and 2R proximal deficiencies associated with the detachment products. Analysis of the 118 EMS-induced recessive lethals and visible mutations recovered provided evidence for seven loci in the 2L heterochromatin and six loci in the 2R heterochromatin, with multiple alleles being obtained for most sites. Of these loci, one in 2L and two in 2R fall near the heterochromatic-euchromatic junctions of 2L and 2R respectively. None of the 113 EMS lethals behaved as a deficiency, implying that the heterochromatic loci uncovered in this study represent nonrepetitive cistrons. Thus functional genetic loci are found in heterochromatin, albeit at a very low density relative to euchromatin.     

Mutagenicity of hycanthone in Drosophila melanogaster

The schistosomicidal agent hycanthone was tested for mutagenicity in Drosophila melanogaster. The compound was administered either by injection into adult males or by larval feeding. The following types of genetic damage were measured:(1) complete and mosaic sex-linked recessive lethal mutations; (2) II–III translocations; and (3) dominant lethals.In postmeiotic germ cells, especially in late spermatids, a pronounced increase was found in the frequency of sex-linked recessive lethals, both completes and mosaics. By contrast, translocations and dominant lethals were not induced.     

Influence of streptomycin on UV-induced TRP+ reversions in a streptomycin-resistant strain of E. Coli B/R

UV mutagenesis has been compared in the E. coli B/r trp^? WWP-2 Hcr⁺ and in a mutant (Stm^R 28) resistant to 100 μg streptomycin/ml. In the Stm^R strain it is possible to score UV-induced Trp⁺ reversions, and survival, on media supplemented with, or lacking, streptomycin. Such experiments revealed a marked enhancement of mutagenic effect of UV by streptomycin. On analysis it was shown that in the Stm^R strain a fraction of UV-induced Trp⁺ reversions, dut to ochre suppressor, achieve full phenotypic expression only in the presence of streptomycin. Thus an inhibitory effect of the Stm^R 28 mutation on the activity of some ochre suppressors is relieved by streptomycin.     

Identification of a gene for alpha-tubulin in Aspergillus nidulans.

This paper demonstrates that revertants of temperature-sensitive benA (β-tubulin) mutations in Aspergillus nidulans can be used to identify proteins which interact with β-tubulin. Three benomyl-resistant benA (β-tubulin) mutants of Aspergillus nidulans, BEN 9, BEN 15 and BEN 19, were found to be temperature-sensitive (ts^?) for growth. Temperature sensitivity co-segregated with benomyl resistance among the progeny of outcrosses of BEN 9, 15 and 19 to a wild-type strain, FGSC#99, indicating that temperature sensitivity was caused by mutations in the benA gene in these strains. Eighteen revertants to ts⁺ were isolated by selection at the restrictive temperature. Four had back-mutations in the benA gene and fourteen carried extragenic suppressor mutations. Two of the back-mutated strains had β-tubulins which differed from the β-tubulins of their parental strains by one (1^?) or two (2^?) negative charges on two-dimensional gel electrophoresis. Although the β-tubulins of the extragenic suppressor strains were all electrophoretically identical to those of the parental strains, one of the suppressor strains, BEN 9R7, had an electrophoretic abnormality in α1-tubulin (1⁺). A heterozygous diploid between this strain and a strain with wild-type α1-tubulin was found to have both wild-type and mutant (1⁺) α1-tubulins. This experiment rules out post-translational modification as a possible cause of the α1-tubulin abnormality. Thus the suppressor mutation in BEN 9R7 must be in a structural gene for α1-tubulin. We propose that this gene be designated tubA to denote that it is a gene for α1-tubulin in A. nidulans.     

A Cytogenetic Analysis of the Chromosomal Region Surrounding the α-Glycerophosphate Dehydrogenase Locus of DROSOPHILA MELANOGASTER

The chromosomal region surrounding the structural gene for α-glycerophosphate dehydrogenase (αGpdh, 2-20.5) of Drosophila melanogaster has been studied in detail. Forty-three EMS-induced recessive lethal mutations and five previously identified visible mutations have been localized within the 25A-27D region of chromosome 2 by deficiency mapping and in some cases by a recombination analysis. The 43 lethal mutations specify 17 lethal loci. αGpdh has been localized to a single polytene chromosome band, 25F5, and there apparently are no lethals that map to the αGpdh locus.     

Mechanism of suppression in Drosophila: a change in tyrosine transfer RNA

The mechanism of suppression of the vermilion locus in Drosophila melanogaster is examined. The suppressor locus, su(s)², is shown to control directly the amount of a specific tyrosine transfer RNA which occurs in the adult fly. Wild-type flies have three chromatographic forms of tyrosine tRNA but flies that are homozygous for the suppressor gene su(s)² contain little or none of the second chromatographic form. The isoacceptor patterns of tRNA for leucine, phenylalanine and serine are identical in the suppressor mutant and wild-type fly. Genetic data show that the phenotypic expression of su(s)² and the altered chromatographic pattern of tyrosine tRNA are recessive and that both map at the same position on the left tip of the X chromosome. Furthermore, another suppressor of vermilion was induced by ethyl methane sulfate, su(s)^e1, that is at the same locus as su(s)² and that produces the same change in tyrosine tRNA as su(s)².     

Keine Mutationsauslösung durch 5-Äthyl-2′-desoxyuridin (ÄDU) bei Drosophila melanogaster

Summary The rate of sex-linked recessive lethals as well as of translocations between 2^nd and 3^rd chromosome was determined in a multipurpose strain ofDrosophila melanogaster after intraperitoneal injection of 5-ethyl-2-deoxyuridine. No mutations, however, were observed after that treatment. These findings confirm the results of other investigators with T-2-phages and are in good agreement with theoretical considerations.     

Radioprotective effect of six chemicals against X-ray-induced genetic damage in D. melanogaster

In Drosophila melanogater six chemicals were tested for radioprotectiveeffect against X-ray-induced genetic damage such as sex-linked recessive lethals and autosomal translocations using Oster's ring-X chromosome stock. A 2-day brood pattern was followed to score the damage induced at different spermatogenic stages separately. In all cases the chemicals were injected before X-irradiation. 10-mM solution of reduced glutathione (GSH) provided statistically significant protection against sex-linked recessive lethals in all broods. In translocation tests this chemical reduced the frequency in all broods but the result is not statistically significant. Cysteamine (MEA) did not show any protective effect but the frequency of lethals was slightly reduced in the first and fourth broods. 2-Aminoethyl isothiuronium Br·HBr (AET) showed a statistically significant protective effect when the data of the replicate experiments were pooled. Negative results were obtained for 5-hydroxytryptamine (5-HT) in sex-linked lethal tests. Aminoethyl phosphorothioate (AEPT) reduced the frequencies of both sex-linked lethals and autosomal translocations in all broods consistently but the results are not statistically significant. In tests for both lethals and translocations the reduction was largest in the stages with highest radiosensitivity. N(3-Aminopropyl)aminoethyl phosphorothioate (3AP-AEPT) gave no protection.     

Chromosomal aberrations induced by caffeine in somatic ganglia of Drosophila melanogaster

Nerve ganglia of third-instar larvae were treated with various doses of caffeine (5×10^?4, 10^?3, 5×10^?3, 10^?2 and 2×10^?2 M) for 2 h at 25±1°C. The ganglia were fixed at set time intervals after treatment so that the effect of caffeine in different stages of the cell cycle could be observed. Chromatid aberrations were induced only when the caffeine was administered in G₂ or approaching mitosis. No aberrations were observed after treatment in S or early G₂. In relation to the different doses administered, a threshold effect was evidenced, the number of aberrations increasing in a marked way at doses exceeding 5×10^?3 M. These data indicate, that the effect observed in Drosophila melanogaster is similar to that described by Kihlman in animals and plants treated with caffeine at temperatures below 30°C.Results obtained in non-cytological tests (non-disjunction, chromosome loss, lethal recessives, dominant lethals) have so far given incomplete indications as to the mutagenicity of caffeine in Drosophila. The results we have obtained with the cytological test seem to contribute to a better definition of the mutagenecity.