Positive correlation between the occurrence of chromosome breakage and the induction of point mutations associated with male recombination 31.1 MRF system of hybrid dysgenesis in Drosophila melanogaster

One of the weak singed (snw) mutations, induced by the 31.1 MRF in the X-chromosome of a laboratory strain, is highly unstable, often changing to either a strong expression (snst) or reverting to wild type (sn+). The present study shows that the X-chromosome carrying the (snw) mutation and the X-chromosome carrying one of the snst alleles derived from the snw mutation generate different frequencies of deletions associated with the w locus. Moreover, they produce different frequencies of mutations associated with the w locus in males after the reintroduction of the 31.1 MRF second chromosome. The occurrence of the deletions and the induction of the mutations are positively correlated and increase when flies are raised at a higher temperature. These data indicate that the induction of the w mutations follows the generation of chromosome breaks in the w locus. The break-points of the recovered deletions occurred in specific sites in the 3C subdivision. Furthermore both snw and snst X-chromosomes induce different frequencies of non-disjunction in females depending on the culture temperature and the genetic background. The present data also show that the 23.5 MRF second chromosome which exhibits specific differences in its activities from the 31.1 MRF is unable to induce w mutations. This fact supports our previous indications that the 31.1 MRF and the 23.5 MRF are not identical.     

Unstable chromosome rearrangements associated with male recombination in Drosophila melanogaster

The male recombination second chromosome 23.5 MRF isolated from the same Greek natural population with the second chromosome 31.1 MRF induced high frequencies of chromosome rearrangements, including specific deletions and duplications. A number of the duplications recovered were found to be highly unstable. The duplicated chromosome segments of the unstable duplications had been either completely or partially lost. The loss occurred most probably by excision of the corresponding segments and not by unequal crossing-over involving sister chromatids. As regards the unstable deletions, they became either shorter or longer or they showed complete restoration. Hypotheses explaining the high frequencies of the unstable chromosome mutations detected are discussed.     

Studies on the chromosomal rearrangements induced by the male recombination factor 31.1 MRF in Drosophila melanogaster

A cytological analysis was carried out on the salivary-gland chromosomes of third-instar larvae deriving from appropriate crosses of the 31.1/CyL⁴ strain with the dp b cn bw and dp b cn bw; ve strains.(1) The 31.1 MRF induced chromosomal rearrangements in both male and female germ cells. (2) The aberrations that occurred in the male germ cells were of three types — inversions, duplications and deficiencies — whereas those that occurred in the female germ cells were of all types. (3) The distributions of the break-points among the 4 major autosomal arms between the 1F₂ and 2AF₂ larvae were different. (4) The total distribution of the break-points along the polytene chromosomes showed that the factor induced breaks in all major chromosomes (X, II, III), and that 51.81% of them occurred in sites showing late replication and/or ectopic pairing. (5) A relationship existed between the frequencies of the second chromosomal aberrations and male recombination.A comparison of the break-points induced by the factor was made with those found in the rare endemic inversions of the same natural population from which the 31.1 MRF was isolated. The possibility that the factor acts in the natural population is discussed.     

hobo is responsible for the induction of hybrid dysgenesis by strains of Drosophila melanogaster bearing the male recombination factor 23.5MRF

The male recombination factor 23.5MRF, isolated ten years ago from a natural Greek population of Drosophila melanogaster, has been shown to induce hybrid dysgenesis when crossed to some M strains, in a fashion slightly different from that of most P strains. Furthermore, it was recently shown that 23.5MRF can also induce GD sterility when crossed to specific P strain females (e.g., Harwich, pi 2 and T-007). In these experiments, the P strains mentioned behaved like M strains in that they did not induce sterility in the reciprocal crosses involving 23.5MRF. We extended the analysis to show that 23.5MRF does not destabilize snW(M) and that a derivative with fewer full-length P elements behaves like an M strain toward the same P strains and still retains its dysgenic properties in the reciprocal crosses. We show that there is a strong correlation between the site of dysgenic chromosomal breakpoints induced by 23.5MRF and the localization of hobo elements on the second chromosome, and also that hobo elements are found associated with several 23.5MRF induced mutations. These results suggest that hobo elements are responsible for the aberrant dysgenic properties of this strain, and that they may express their dysgenic properties independent of the presence of P elements.     

Chromosome interactions in P-M dysgenic male recombination of Drosophila melanogaster.

The frequency, distribution and structure of P elements on the second and third chromosomes of Texas 1, a wild-type inbred strain of Drosophila melanogaster, were investigated by in situ hybridization. These autosomes were isolated individually and used as P-element donors to study the frequency and distribution of male recombination events generated on recipient chromosomes which were originally devoid of P sequences. The P-element array of chromosome 2 was shown to generate higher male recombination frequencies on chromosome 3 than vice versa, despite having fewer P factors and fewer P elements in general. This is likely to be due to the presence and distribution of specific P-deletion derivatives, which vary in their ability to repress P mobility. The male recombination generated on recipient chromosomes is associated with the insertion of donated P sequences, but only in a small minority of cases could a novel P-element site be detected at, or near, the recombination breakpoint. The majority of such breakpoints appear to be associated either with unsuccessful P insertion, or with the action of P transposase attracted by P elements newly inserted elsewhere on the recipient chromosome. Recent evidence also suggests that a small proportion of the breakpoints may be associated with the action of P transposase alone. Male recombination breakpoints appear to be distributed effectively at random along the recipient autosomes, and their frequency of occurrence was shown to correlate with the physical length of DNA available between markers, as revealed by the polytene map distance.     

Characterization of 5' truncated transposed copies of the I factor in Drosophila melanogaster.

I factors in Drosophila melanogaster are transposable elements structurally related to Mammalian LINEs. Their transposition is activated at high frequencies during I-R hybrid dysgenesis and is associated with the production of mutations of various sorts. Very few of these mutations have been studied at the molecular level; those reported so far result either from chromosomal rearrangements or from insertions of complete I factors. We have analysed three I-R induced yellow mutations and have found that one of them is due to the insertion of an I element very similar to the complete I factor, whereas the other two are due to insertions of I elements that are truncated at their 5' ends; one of them exhibits an unusual 3' end. We discuss possible mechanisms of production of such modified I elements.     

Analysis of male recombination in third chromosomes of Drosophila melanogaster

Data on male recombination in twenty third-chromosomal lines of Drosophila melanogaster are presented. Frequencies of female and male recombination have been calculated in seven intervals along the third chromosome. The influence on male recombination (M.R.) exercised by different factors such as population origin (cellar, vineyard), the presence of heterozygous inversions and recessive lethal chromosomes, is analyzed. The results obtained lead to the main conclusion that M.R. is not affected by the presence of heterozygous inversions which reduce female recombination in the same lines. In the light of this effect, the possible mechanism operating on male recombination is discussed. Lethal chromosomes reduce significantly the number of male recombination events as compared with wild chromosomes. We have not obtained significant differences in male recombination frequencies between the cellar and the vineyard lines.     

Induction of male recombination in Drosophila melanogaster by chemical treatment

Acridine orange (AO), ethidium bromide (EB), ethyl methanesulfonate (EMS) and 8- ethoxycaffeine ( EOC ) were fed to larvae of Drosophila melanogaster in order to test their capacity for the induction of meiotic recombination in males. Our results show that AO and EB increase significantly the male recombination frequencies. No relationship between chromosome breakage ability and male recombination induction was found since EMS and EOC , two effective chromosome-breaking agents, were unable to increase the male recombination.     

Time of recombination in the Drosophila melanogaster Oocyte

A test has been carried out to determine if the restrictive temperature (31°) acts to reduce recombination in the temperature-sensitive recombination-deficient genotype rec-1 ²⁶/rec-1 ¹⁶ by reducing or eliminating the synaptonemal complex. Measurements of the length of synaptonemal complexes in heat-treated and untreated stage 1 oocytes, following termination of the temperature-sensitive period, reveal less than a 5% difference, with the greater length present in the treated oocytes. Alterations are not observed in synaptonemal complex distribution within the nucleus or in its fine structure. Parallel genetic studies confirm earlier observations that the restrictive temperature, whose action is confined to a 36-h sensitive period virtually coextensive with premeiotic-S, drastically reduces recombination to 10% of normal. The results are most simply interpreted to mean that the restrictive temperatures acts directly on the recombination process.     

Asymmetric exchange is associated with P element induced male recombination in Drosophila melanogaster

Spontaneous meiotic recombination events do not normally occur in the male germ line of Drosophila melanogaster. However, such events are induced in males when a P transposable element or a source of P element encoded transposase protein is present in its genome. This report concerns a molecular analysis of the meiotic exchanges that were induced in the male Drosophila by P elements within a genetically marked region of the third chromosome. The marked region also harbors a single P-element called P(lArB). Fifty-six percent of the P(lArB) region crossovers indicated some alterations in the P element 5' fragment. Such alterations appear to be related to asymmetric or unequal genetic exchanges. Finally, P(lArB) excision was found to be independent of P(lArB) region crossover events.