Analysis of Y-Linked Mutations to Male Sterility in DROSOPHILA MELANOGASTER

Mating type in haploid cells of the yeast Saccharomyces cerevisiae is determined by a pair of alleles MATa and MAT alpha. Under various conditions haploid mating types can be interconverted. It has been proposed that transpositions of silent cassettes of mating-type information from HML OR HMR to MAT are the source of mating type conversions. A mutation described in this work, designated AON1, has the following properties. (1) MAT alpha cells carring AON1 are defective in mating. (2) AON1 allows MAT alpha/MAT alpha but not MATa/MATa diploids to sporulate; thus, AON1 mimics the MATa requirement for sporulation. (3) mata-1 cells that carry AON1 are MATa phenocopies, i.e., MAT alpha/mata-1 AON1 diploids behave as standard MAT alpha/MATa cells; therefore, AON1 suppresses the defect of mata-1. (4) AON1 maps at or near HMRa. (5) Same-site revertants from AON1 lose the ability to convert mating type to MATa, indicating that reversion is associated with the loss of a functional HMRa locus. In addition, AON1 is a dominant mutation. We conclude that AON1 is a regulatory mutation, probably cis-acting, that leads to the constitutive expression of silent a mating-type information located at HMRa.     

Male-Specific Lethal Mutations of DROSOPHILA MELANOGASTER

A total of 7,416 ethyl methanesulfonate (EMS)-treated second chromosomes and 6,212 EMS-treated third chromosomes were screened for sex-specific lethals. Four new recessive male-specific lethal mutations were recovered. When in homozygous condition, each of these mutations kills males during the late larval or early pupal stages, but has no detectable effect in females. One mutant, mle^ts, is a temperature sensitive allele of maleless, mle (Fukunaga, Tanaka and Oishi 1975), while the other three mutants identify two new loci: male-specific lethal-1 (msl-1) (two alleles) at map position 2-53.3 and male-specific lethal-2 (msl-2) at 2-9.0.——The male-specific lethality associated with these mutants is not related to the sex per se of the mutant flies, since sex-transforming genes fail to interact with these mutations. Moreover, the presence or absence of a Y chromosome in males or females has no influence on the male-specific lethal action of these mutations. Finally, no single region of the X chromosome, when present as a duplication, is sufficient to rescue males from the lethal effects of msl-1 or msl-2. These results suggest that the number of complete X chromosomes determines whether a fly homozygous for a male-specific lethal mutation lives or dies.     

Selection for Male Recombination in DROSOPHILA MELANOGASTER

Two-way selection for male recombination over seven intervals of the third chromosome in Drosophila melanogaster was practiced for nine generations followed by relaxed selection for five generations. Significant responses in both directions were observed but these mainly occurred in early generations in the low line and in later generations in the high line. Divergence of male recombination frequencies between the two selection lines was not restricted to any specific region but occurred in every measured interval of the chromosome. However, right-arm intervals showed a more pronounced response than either left-arm intervals or the centromeric region. Correlated responses in sterility and distortion of transmission ratios occurred as a result of selection for male recombination. Cluster distributions of male recombinants suggested a mixture of meiotic and late gonial events but relative map distances more closely resembled those of the salivary chromosome than standard meiotic or mitotic distances. Patterns of male recombination over time in both second and third chromosomes strongly suggested a major effect associated with the presence of third chromosomes from the Harwich strain. Evidence was also found for modifiers with relatively small effects located in other regions of the genome. The overall results are interpreted in terms of a two-component model of hybrid dysgenesis.     

High Mutability in Male Hybrids of DROSOPHILA MELANOGASTER

The frequencies of sex-linked lethal mutations arising in hybrid male offspring from various crosses and in nonhybrid controls were determined. The hybrids were produced by crossing representative strains of the P-M system of hybrid dysgenesis in all possible combinations. Males from the cross of P males x M females had a mutation rate about 15 times higher than that of nonhybrid males from the P strain. Genetically identical males from the reciprocal cross had a mutation rate 3 to 4 times that of the nonhybrids. For crosses involving a Q strain, a significant increase in the mutation rate was detected in males produced by matings of Q males with M females. No increase was observed in genetically identical males from the reciprocal mating. Crosses between P and Q strains gave male hybrids with mutation rates not different from those of nonhybrids. Many of the lethals that occurred in hybrids from the cross of P males x M females appeared to be unstable; fewer lethals that arose in hybrids from the cross of Q males x M females were unstable. The relationship between P and Q strains is discussed with respect to a model of mutation induction in dysgenic hybrids.     

Density-Dependent Fertility Selection in Experimental Populations of DROSOPHILA MELANOGASTER

The effects of larval density on components of fertility fitness were investigated with two mutant lines of Drosophila melanogaster. The differences in adult body weight, wing length, larval survivorship and development time verified that flies reared at high density were resource limited. Experimental results indicate that: (1) relative fecundities of both sexes show density-dependent effects, (2) there is a strong density effect on male and female mating success, and (3) in general, there is a reduction in fecundity differences between genotypes at high density. These results imply that it may be important to consider fertility in models of density-dependent natural selection.     

Elements Causing Male Crossing over in DROSOPHILA MELANOGASTER

A second chromosome line of Drosophila melanogaster (Symbol: T-007) has previously been shown to be responsible for the induction of male recombination. In the present investigation, the genetic elements responsible for this phenomenon have been partially identified and mapped. A major element (Symbol: Mr, for Male recombination) locates on the second chromosome between the pr (2L-54.4) and c (2R-75.5) loci and is responsible for the large majority of male recombination. In addition, there appear to be "secondary elements" present which have the ability to induce male recombination in much reduced frequencies and which are diluted out through successive backcross generations when Mr is removed by recombination. The possible nature of these "secondary elements" is discussed.     

Multiple Meiotic Drive Systems in the DROSOPHILA MELANOGASTER Male

The behaviour of two "meiotic drive" systems, Segregation-Distorter (SD) and the sex chromosome sc(4)sc(8) has been examined in the same meiocyte. It has been found that the two systems interact in a specific way. When the distorting effects of SD and sc(4)sc(8) are against each other, there is no detectable interaction. Each system is apparently oblivious to the presence of the other, gametes being produced according to independence expectations. However when the affected chromosomes are at the same meiotic pole an interaction occurs; the survival probability of the gamete containing both distorted chromosomal products is increased, rather than being decreased by the combined action of two systems.     

Mutations in Genes Encoding Essential Mitotic Functions in DROSOPHILA MELANOGASTER

Temperature-sensitive mutations at 15 loci that affect the fidelity of mitotic chromosome behavior have been isolated in Drosophila melanogaster. These mitotic mutants were detected in a collection of 168 EMS-induced X-linked temperature-sensitive (ts) lethal and semilethal mutants. Our screen for mutations with mitotic effects was based upon the reasoning that under semirestrictive conditions such mutations could cause an elevated frequency of mitotic chromosome misbehavior and that such events would be detectable with somatic cell genetic techniques. Males hemizygous for each ts lethal and heterozygous for the recessive autosomal cell marker mwh were reared under semirestrictive conditions, and the wings of those individuals surviving to adulthood were examined for an increased frequency of mwh clones. Those mutations producing elevated levels of chromosome instability during growth of the wing imaginal disc were also examined for their effects on chromosome behavior in the cell lineages producing the abdominal cuticle. Fifteen mutations affect chromosome behavior in both wing and abdominal cells and thus identify loci generally required for the fidelity of mitotic chromosome transmission. Mapping and complementation tests show that these mutations represent 15 loci. One mutant is an allele of a locus (mus-101) previously identified by mutagen-sensitive mutants and a second mutant is an allele of the lethal locus zw 10.--The 15 mutants were also examined cytologically for their effects on chromosomes in larval neuroblasts. Taken together, the results of our cytological and genetical studies show that these mutants identify loci with wild-type functions necessary for either maintenance of chromosome integrity or regular disjunction of chromosomes or chromosome condensation. Thus, these mutations define a broad spectrum of genes required for the normal execution of the mitotic chromosome cycle.     

Clonal Analysis of Dominant Female-Sterile, Germline-Dependent Mutations in DROSOPHILA MELANOGASTER

Three allelic, dominant and germline-dependent female-sterile mutations (ovo(D) mutations) can be classified according to the severity of the ovarian abnormalities that they produce. The size and frequency of +/+ germline clones, induced in ovo(D)/+ females, were compared with K10/K10 germline clones induced in K10/+ control females. The frequency of germline clones induced by irradiation of first instar larvae is similar for the three dominant alleles and K10 ; however, the clone size increased with the strength of the allele tested, compared with K10 clones. When clones were induced later in development, the clone frequencies decreased with the strength of the alleles. These results are discussed in the context of the antimorphic nature of these mutations and the characteristics of germline development. The use of these alleles as tools in the genetic analysis of development is discussed.     

Investigation of the Nature of P-Induced Male Recombination in DROSOPHILA MELANOGASTER

The present study consists of an investigation of P-induced male recombination in Drosophila melanogaster from a number of perspectives. In an initial set of experiments, male recombination induced by several different P strains was examined on both major autosomes. The ability of these P strains to evoke recombination is striking; in many cases it exceeded that of radiation treatment. Also of interest is the apparent nonrandom chromosomal distribution of P-exchange breakpoints. The data suggest that both recombinagenic capacity and distribution pattern of exchange breakpoints may be P-strain specific. In addition to these findings, we have confirmed previous indications that P-induced exchange is reasonably symmetrical and that it frequently occurs during premeiotic stages of spermatogenesis. Moreover, we have established that radiation and P background act additively with regard to the induction of male recombination. The second part of the work involved an analysis of heterochromatic vs. euchromatic recombination induced by several recombinagenically potent P strains. Results of these experiments have confirmed our earlier findings concerning the recombinagenic capacity of p strains. More importantly, it would appear that P-induced exchange in heterochromatin is rare. The induction of various kinds of mutations was also monitored in several of these experiments. The results indicate that the mutagenic potential of the P strains is substantial and of particular interest, that certain types of mutations are P-strain specific. For example, rare heterochromatic lesions were recovered exclusively in the experiment using the h12 strain, whereas a novel pleiotropic mutation occurred at a high frequency only in the T-007 experiment. Our findings are discussed within the context of a model of P-induced exchange.     

Hybrid Dysgenesis in DROSOPHILA MELANOGASTER: The Biology of Female and Male Sterility

High levels of female and male sterility were observed among the hybrids from one of the two reciprocal crosses between a wild strain of D. melanogaster known as pi2 and laboratory strains. The sterility, which is part of a common syndrome called hybrid dysgenesis, was found to be associated with the rudimentary condition of one or both of the ovaries or testes. All other tissues, including those of the reproductive system were normal, as were longevity and mating behavior. The morphological details of the sterility closely mimic the agametic condition occurring when germ cells are destroyed by irradiation or by the maternal-effect mutation, grandchildless. We suggest that sterility in hybrid dysgenesis is also caused by failure in the early development of germ cells. There is a thermo-sensitive period beginning at approximately the time of initiation of mitosis among primordial germ cells a few hours before the egg hatches and ending during the early larval stages. Our results suggest that hybrid dysgenesis, which also includes male recombination, mutation and other traits, may be limited to the germ line, and that each of the primordial germ cells develops, or fails to develop, independently of the others. This hypothesis is consistent with the observed frequencies of unilateral and bilateral sterility, with the shape of the thermosensitivity curves and with the fact that males are less often sterile than females. The features of this intraspecific hybrid sterility are found to resemble those seen in some interspecific Drosophila hybrids, especially those from the cross D. melanogaster X D. simulans.     

Spontaneous Chromosome Breakage at Male Meiosis Associated with Male Recombination in DROSOPHILA MELANOGASTER

An inbred line (OK1) of Drosophila melanogaster , recently derived from a natural population in Oklahoma, has been found by Woodruff and Thompson to exhibit a low frequency of spontaneous male recombination when outcrossed to marker stocks. There is also a reciprocal-cross effect, such that recombination is found only if OK1 males are used in the initial cross. When OK1 females are used, however, male recombination is again found if their male progeny are used for a subsequent cross.-In the present cytological analysis, chromosome behavior at male meiosis was studied in reciprocal crosses between the OK1 line and both a marker gene stock and an inversion stock. If the recombination events were "conventional" and premeiotic (gonial) in origin, no chromosome aberrations would be expected during meiosis. If they were "conventional" and meiotic, some dicentric bridges with free fragments would be expected in the inversion heterozygote, but none should be present in the marker gene cross.-The results demonstrated that the occurrence of recombination in males is most likely a meiotic event, though the occurrence of some limited premeiotic recombination can not be disproven. Meiosis was found to be perfectly normal in all crosses lacking male recombination. In all of the inversion stock and noninversion marker stock crosses that showed male recombination, however, anaphase bridges were found at both first and second meiotic divisions. These were often accompanied by more than the single fragment expected from a conventional inversion bridge and fragment situation. In extreme cases, almost complete pulverization of one or more autosomes was found.-All metaphase I stages were perfectly normal, suggesting that no comparable breakage occurs in premeiotic gonial mitoses. The form of chromosome damage is similar in many ways to that produced by some DNA synthesis inhibitors, or by some viral or mycoplasma infections. This possibility is discussed, and some of the evolutionary implications of the system are briefly considered.