Super-unstable mutations associated with P-M hybrid dysgenesis in Drosophila melanogaster

Super-unstable mutations occasionally appear either in natural populations of Drosophila melanogaster or in P-M hybrid dysgenesis. We found that they may be reproducibly obtained with a high frequency from crosses between males from the % MathType!MTEF!2!1!+-% feaafeart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGqiVu0Je9sqqrpepC0xbbL8F4rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaWaaubiaeqale% qabaWaaubiaeqameqaleaacaGGQaaaoeaacaWGHbGaam4raaaaa0qa% aiaadEhaaaaaaa!3A01!\[\mathop w\nolimits^{\mathop {aG}\nolimits^* } \] strain and females from the w ^aG* strain or its derivatives. Super-unstable mutations in the ocelliless, singed, white, yellow and other loci have been obtained. Each super-unstable mutation gives rise to a large family of new super-unstable mutations with a wide range of phenotypic expression. Mutations with the same phenotype often differ in the specificity of their potential for further mutation. As a rule, a super-unstable mutation is associated with a specific reversible mutation and paired alleles are formed in this way. Other mutations are usually irreversible, but new mutations of these may also form paired alleles. Active transposase encoded by transposable P elements is necessary to maintain super-instability. Finally, some preliminary molecular data are discussed which suggest that this type of super-instability is a result of interaction between P elements and a novel mobile element, designated as X.     

P-M hybrid dysgenesis affects juvenile hormone metabolism in Drosophila melanogaster females

This paper studies the metabolism of the juvenile hormone, which affects gonads functioning in Drosophila melanogaster females under P-M hybrid dysgenesis. It is shown that dysgenic females grown at 29°C have increased levels of the juvenile hormone (its degradation and stress reactivity are reduced), which apparently is a compensatory response to ovarian hypoplasia.     

Genetic and molecular analysis of repression in the P-M system of hybrid dysgenesis in Drosophila melanogaster.

Twelve inbred lines derived from an M' strain of Drosophila melanogaster were used to study the repression of P-element-mediated hybrid dysgenesis. Initial assessments indicated that the lines differed in the ability to repress gonadal dysgenesis, and that this ability was highly correlated with the ability to repress snw hypermutability. Later assessments indicated that most of the lines with low or intermediate repression potential evolved to a state of higher repression potential; however, Southern analyses failed to reveal significant changes in the array of genomic P elements that could account for this evolution. In addition, none of the lines possessed the incomplete P element known as KP, which has been proposed to explain repression in some D. melanogaster strains. One of the lines maintained intermediate repression potential throughout the period of study (52 generations), indicating that the intermediate condition was not intrinsically unstable. Genetic analyses demonstrated that in some of the lines, repression potential was influenced by factors that were inherited maternally through at least two generations; however, these factors were not as influential as those in a classic P cytotype strain. Additional tests with a dysgenesis-inducing X chromosome called T-5 indicated that repression itself was mediated by a combination of maternal effects and paternally inherited factors that were expressed after fertilization. These tests also suggested that in some circumstances, the P transposase, or its message, might be transmitted through the maternal cytoplasm.     

Visible mutations induced by P-M hybrid dysgenesis in Drosophila melanogaster result predominantly from P element insertions.

This study supplied no evidence that P-M hybrid dysgenesis is a general release mechanisms for transposon movement. Newly induced mutations (23 singed, three yellow, and one white) were generated by hybrid dysgenesis and were molecularly analyzed for the presence or absence of P element insertions. Only one dysgenically-induced insertion mutation out of 27 analyzed was the result of a non-P insert; this frequency is not statistically above the non-dysgenic control level. Thus, it appears that individual transposable elements families are independently regulated.     

KP elements repress P-induced hybrid dysgenesis in Drosophila melanogaster.

Molecular and genetic analysis has revealed a specific P factor deletion derivative (the KP element) which is able to repress P-induced hybrid dysgenesis. All naturally occurring strains lacking the P cytotype (M') that were examined, throughout the world contain up to 30 copies of KP per haploid genome together with complete P factors. The KP element is derived from the P factor by an internal deletion of 1753 bp removing nucleotides 808-2560 and is transcribed to yield an abundant 0.8-kb poly(A)+ RNA with the coding capacity for an in-frame 207 amino acid polypeptide. Genetic crosses show that KP elements preferentially accumulate in the presence of P factors and suppress hybrid dysgenesis. Suppression is transmitted through both sexes and is thus distinct from the maternally transmitted P cytotype mode of suppression. The spread of KP elements is probably due to the continual selection of individuals with the highest numbers of KP elements in which P-induced hybrid dysgenesis is suppressed.     

Hybrid dysgenesis in Drosophila melanogaster: influence of temperature on cytotype determination in the P-M system

Summary In Drosophila melanogaster, the P-M system of hybrid dysgenesis is a syndrome of germ line abnormalities, including temperature dependent gonadal dysgenesis (GD sterility), high rates of mutation and male recombination, which occurs in some interstrain hybrids but only from one of the two crosses. In the P-M system, hybrid dysgenesis results from interaction between chromosomally transposable elements of the P element family and a particular extrachromosomal state referred to as the M cytotype. Cytotype (M or P) is known to be determined by the absence or presence of chromosomal factors, but principally with limited cytoplasmic transmission.In a series of experiments in which F1 hybrid females from various P and M strains were submitted to different preadult and ageing temperature treatments, it was found that the cytotype switch is strongly temperature-dependent in the F1 females from M x P but not in the reciprocal cross. In the F1 females from the former cross, a strong M cytotype occurs at a low developmental temperature (18° C) and a weak M cytotype occurs at a high developmental temperature (26.5° C). On the other hand, a high ageing temperature applied after a low developmental temperature switches the cytotype from M to P and reciprocally, a low ageing temperature applied after a high developmental temperature switches the cytotype from P to M.This thermo-reversibility of the extrachromosomal state exists only in the F1 females from M mothers but not in the F1 females from P mothers; this dissymmetrical behavior is discussed in relation to the mechanism proposed by O'Hare and Rubin (1983) which explains cytotype determination by a positive feedback of the regulator of the P transposase on its own level of activity.     

Evolutionary flux of P element regulation in a Drosophila melanogaster hybrid dysgenesis cline.

Clines of P-induced hybrid dysgenesis provide a means for monitoring the evolution of transposition repression over space and time. We have studied the molecular and phenotypic profiles of flies taken from a 2900 km cline along the eastern coast of Australia, which had previously been characterized over 10 years ago as having P populations in the north, Q populations at central sites and M' populations in the south. We have found that Q and M' populations of flies have increased their range within the cline at the expense of P lines. Q populations were found to be in the north of the cline and M' populations in the south. Some of the northern Q lines transmit repression through both sexes and type I deletion elements have been isolated from them. We suggest that these elements are responsible for Q type repression. The results support our model that populations made up of Q individuals with strong biparentally transmitted repression form an evolutionarily stable strategy for the repression of hybrid dysgenesis in Drosophila melanogaster.     

Characteristic features of induced mutagenesis in hybrid dysgenesis systems of Drosophila melanogaster

The mutagenic effect of low-dose gamma-irradiation was studied in Drosophila melanogaster systems of hybrid dysgenesis by estimating polytene chromosome rearrangements, recombination frequency, and viability at the embryonic and postembryonic developmental stages. A dose of gamma-irradiation which had no effect detectable by routine line crossing proved to significantly reduce the number of recombinants in the H-E and P-M systems and mortality at postembryonic stages. However, this combined effect was obtained if irradiation followed transposition, i.e., it depended on the application sequence of the mutagenic factors. The reverse order of the mutagenic treatment led to summation of the effects: as compared to either control, the frequencies of the dominant allele mutations as well as the larval and pupal mortality in F2 increased significantly (at the level of 99.9%). This allowed us to estimate the contribution of extremely low-dose gamma-irradiation into the mutagenic effect, which was impossible under routine conditions.     

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.     

Structure and genomic organization of I elements involved in I-R hybrid dysgenesis in Drosophila melanogaster.

I-R hybrid dysgenesis in D. melanogaster is controlled by transposable elements known as I factors which terminate at their 3' ends by an A-rich sequence. Inducer strains contain active I factors. Both reactive and inducer stocks possess defective I elements. We have cloned various I elements from both categories of strains. The I elements having recently transposed in inducer strains have a structure closely related to that of active I factors. However we have isolated one such I element that is truncated at its 5' end. The I elements common to reactive and inducer strains are affected by various rearrangements and many point mutations. They do not appear to be simple derivatives of complete I factors.     

Progressive loss of DNA sequences from terminal chromosome deficiencies in Drosophila melanogaster.

Terminal deficiencies at the tip of the X chromosome can be induced at a high frequency (0.2-0.3%) by irradiating Drosophila females carrying a homozygous mutator (mu-2) with low doses of X-rays. These terminal deficiencies are unstable, since over a period of 3 1/2 years DNA sequences were lost from their distal ends at a rate of 75 bp per generation, presumably due to the absence of a complete wild-type telomeric structure. Breakpoints of these deletions in the 5' upstream regulatory region of the yellow gene, giving rise to a mosaic cuticle pigmentation pattern typical of the y2 type, were used to define the location of tissue-specific cis-acting regulatory elements that are required for body, wing or bristle pigmentation.     

Enzymatic studies of DNA repair in Drosophila melanogaster

Thus far, our studies in Drosophila have concentrated primarily on the various enzymes involved in the in vitro repair of modified or nonconventional DNA substrates. In some cases, our findings have led us to investigate events that may not have a bearing on DNA repair, but rather may be associated with developmental signals important to the maturation of the organism. As appealing as some of these models seem, however, they must await confirmation through detailed genetic studies before any substantial conclusions can be drawn. This combination of genetic and biochemical knowledge makes Drosophila an exciting organism for an eventual detailed understanding of the developmental expression and cellular location of DNA-repair systems.