Evidence for rapid limitation of the I element copy number in a genome submitted to several generations of I-R hybrid dysgenesis in Drosophila melanogaster

Summary When Drosophila melanogaster males coming from a class of strains known as inducer are crossed with females from the complementary class (reactive), a quite specific kind of sterility is observed in the F₁ female progeny (denoted SF). The inducer chromosomes differ from the reactive chromosomes by the presence of a transposable element (called the I factor) that is responsible for the induction of this dysgenic symptom. In the germ line of dysgenic females, up to 100% of the reactive chromosomes may be contaminated, i.e. they acquire I factor(s) owing to very frequent replicative transpositions. A contaminated reactive stock was obtained by reconstructing the reactive genotype in the offspring of SF females and its kinetics of invasion by I elements was followed in the successive inbred dysgenic generations. The results show that the mean copy number of I elements increased very quickly up to the level of inducer strains and then stayed in equilibrium even though the dysgenic state was perpetuated by selection for SF sterility at every generation. The possible mechanisms of this copy number limitation are discussed.     

Defective I elements introduced intoDrosophila as transgenes can regulate reactivity and prevent I-R hybrid dysgenesis

The I-R hybrid dysgenesis syndrome is characterized by a high level of sterility and I element transposition, occurring in the female offspring of crosses between males of inducer (I) strains, which contain full-length transposable I elements, and females of reactive (R) strains, devoid of functional I elements. The intensity of the syndrome in the dysgenic cross is essentially dependent on the reactivity level of the R females, which is ultimately controlled by still unresolved polygenic chromosomal determinants. In the work reported here, we have introduced a transposition-defective I element with a 2.6 kb deletion within its second open reading frame into a highly reactive R strain, by P-mediated transgenesis. We demonstrate that this defective I element gradually alters the level of reactivity in the three independent transgenic lines that were obtained, over several generations. After > 15 generations, the transgenicDrosophila show strongly reduced reactivity, and finally become refractory to hybrid dysgenesis, without, however, acquiring the inducer phenotype. Induction of a low reactivity level is reversible reactivity again increases upon transgene removal and is maternally inherited, as observed for the control of reactivity in natural R strains. These results demonstrate that defective I elements introduced as single-copy transgenes can act as regulators of reactivity, and suggest that some of the ancestral defective pericentromeric I elements that can be found in all reactive strains could be the molecular determinants of reactivity.     

Control of expression of the I factor, a LINE-like transposable element in Drosophila melanogaster.

I factors are LINE-like transposable elements in the genome of Drosophila melanogaster. They normally transpose infrequently but are activated in the germline of female progeny of crosses between males of a strain that contains complete elements, an I or inducer strain and females of a strain that does not, an R or reactive strain. This causes a phenomenon known as I-R hybrid dysgenesis. We have previously shown that the I factor promoter lies between nucleotides 1 and 30. Here we demonstrate that expression of this promoter is regulated by nucleotides 41-186 of the I factor. This sequence can act as an enhancer as it stimulates expression of the hsp7O promoter in ovaries in the absence of heat-shock. Within this region there is a site that is required for promoter activity and that is recognized by a sequence-specific binding protein. We propose that this protein contributes to the enhancer activity of nucleotides 41-186 and that reduced I factor expression in inducer strains is due to titration of this protein or others that interact with it.     

Defective I elements introduced intoDrosophila as transgenes can regulate reactivity and prevent I-R hybrid dysgenesis

The I-R hybrid dysgenesis syndrome is characterized by a high level of sterility and I element transposition, occurring in the female offspring of crosses between males of inducer (I) strains, which contain full-length transposable I elements, and females of reactive (R) strains, devoid of functional I elements. The intensity of the syndrome in the dysgenic cross is essentially dependent on the reactivity level of the R females, which is ultimately controlled by still unresolved polygenic chromosomal determinants. In the work reported here, we have introduced a transposition-defective I element with a 2.6 kb deletion within its second open reading frame into a highly reactive R strain, by P-mediated transgenesis. We demonstrate that this defective I element gradually alters the level of reactivity in the three independent transgenic lines that were obtained, over several generations. After > 15 generations, the transgenicDrosophila show strongly reduced reactivity, and finally become refractory to hybrid dysgenesis, without, however, acquiring the inducer phenotype. Induction of a low reactivity level is reversible reactivity again increases upon transgene removal and is maternally inherited, as observed for the control of reactivity in natural R strains. These results demonstrate that defective I elements introduced as single-copy transgenes can act as regulators of reactivity, and suggest that some of the ancestral defective pericentromeric I elements that can be found in all reactive strains could be the molecular determinants of reactivity.     

Germ-line expression of the I factor, a functional LINE from the fruit fly Drosophila melanogaster, is positively regulated by reactivity, a peculiar cellular state

I factor is a functional LINE (long interspersed nucleotidic element) which is mobilized in the germ-line of dysgenic SF females during I-R hybrid dysgenesis. Such females are obtained when an oocyte from a reactive stock, devoid of I factors but characterized by a level of reactivity, i.e. its potential for hybrid dysgenesis, is fertilized by a spermatozoon from an I factor-containing inducer stock. In a previous paper we described the expression of an I factor-lacZ fusion. Expression was detected in the ovaries of reactive and dysgenic flies only. In this paper we show that this transgenic activity can be quantified and depends upon the maternally inherited reactivity. Reactivity is not just a permissive state and modifiers of the reactivity level such as heat treatment and ageing change the level of expression of our transgenic fusion accordingly. Moreover, ageing through generations has the same cumulative and reversible effect on both reactivity and I factor expression. Using our fusion as a test for reactivity we show that the silencing of I factor after its introduction into a reactive genome may not be established in a single generation.     

The I-R system of hybrid dysgenesis in Drosophila melanogaster: influence on SF females sterility of their inducer and reactive paternal chromosomes.

A specific kind of sterile F1 female, denoted SF, arises when females from strains known as reactive are crossed with males from the complementary class of strains (inducer). It has been shown that this sterility results from the interaction between the maternal reactive cytoplasm and any one of the paternal inducer chromosomes. This interaction yields other dysgenic traits including non-disjunction and mutations. In this note, the abilities of paternal gametes containing various combinations of inducer and reactive chromosomes to give more or less sterile SF females when fertilising standard reactive oocytes were compared. Although they did not cause SF sterility, reactive chromosomes, when present in sperm containing at least one inducer chromosome, were found to influence the intensity of sterility: variations of SF sterility were observed between SF females which differed only by one paternally inherited reactive chromosome. Reactive chromosomes are known to control the cytoplasmic state of reactive females. The present results suggest that this chromosomal control also takes place in SF females.     

Non-Mendelian Female Sterility in DROSOPHILA MELANOGASTER: Characterization of the Noninducer Chromosomes of Inducer Strains

In relation to non-Mendelian female sterility, Drosophila melanogaster strains can be divided into two main classes, inducer and reactive. The genetic element responsible for the inducer condition (I factor) is chromosomal and may be linked to any inducer-strain chromosome. Each chromosome carrying the I factor (i(+) chromosome) can, when introduced by the paternal gamete into a reactive oocyte, give rise to females (denoted SF) showing more-or-less reduced fertility. As long as i(+) chromosomes are transmitted through heterozygous males with reactive originating chromosomes (r chromosomes), I factor follows Mendelian segregation patterns. In contrast, in heterozygous i(+)/r females, a varying proportion of r chromosomes may irreversibly acquire I factor, independently of classical genetic recombination, by a process called chromosomal contamination. The contaminated reactive chromosomes behave as i(+) chromosomes.-In the present paper, evidence is given that the Luminy inducer strain displays a polymorphism for two kinds of second chromosomes. Some of them are i(+), while others, denoted i(o), are unable to induce any SF sterility when introduced by paternal gametes into reactive oocytes. They are also unable to induce contamination of r chromosomes, but, like r chromosomes, they may be contaminated by i(+) chromosomes in SF or RSF females. The study of the segregation of i(+) and i(o) second chromosomes in the progeny of heterozygous Luminy males and females leads to the conclusion that on chromosome 2 of the Luminy stock the I factor is at a single locus. -X, second and third i(o) chromosomes have been found in several inducer strains. Since these chromosomes can be maintained with i(+) chromosomes in inducer strains in spite of their ability to be contaminated in RSF females, it can be concluded that chromosomal contamination does not take place in females of inducer strains. This implies that contamination occurs only in cells having cytoplasm in a reactive state.     

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.     

Molecular characteristics of the heterochromatic I elements from a reactive strain ofDrosophila melanogaster

Summary There are two categories of strains inDrosophila melanogaster with respect to the I-R system of hybrid dysgenesis. The inducer strains contain particular transposable elements named I factors. They are not present in the strains of the other category called reactive (R) strains. Defective I elements are present in the pericentromeric regions of both categories of strains. This last subfamily of I sequences has not yet been described in detail and little is known about its origin. In this paper, we report that the defective I elements display an average of 94% of sequence identity with each other and with the transposable I factor. The results suggest that they cannot be the progenitors of the present day I factors, but that each of these two subfamilies started to evolve independently several million years ago. Furthermore, the sequence comparison of these I elements with an active I factor fromDrosophila teissieri provides useful information about when the deleted I elements became immobilized.     

Germ-line expression of the I factor,a functional LINE from the fruit fly Drosophila melanogaster,is positively regulated by reactivity,a peculiar cellular state

I factor is a functional LINE (long interspersed nucleotidic element) which is mobilized in the germ-line of dysgenic SF females during I-R hybrid dysgenesis. Such females are obtained when an oocyte from a reactive stock, devoid of I factors but characterized by a level of reactivity, i.e. its potential for hybrid dysgenesis, is fertilized by a spermatozoon from an I factor-containing inducer stock. In a previous paper we described the expression of an I factor-lacZ fusion. Expression was detected in the ovaries of reactive and dysgenic flies only. In this paper we show that this transgenic activity can be quantified and depends upon the maternally inherited reactivity. Reactivity is not just a permissive state and modifiers of the reactivity level such as heat treatment and ageing change the level of expression of our transgenic fusion accordingly. Moreover, ageing through generations has the same cumulative and reversible effect on both reactivity and I factor expression. Using our fusion as a test for reactivity we show that the silencing of I factor after its introduction into a reactive genome may not be established in a single generation.     

Non-Mendelian female sterility in Drosophila melanogaster: influence of ageing and thermic treatments. I. Evidence for a partly inheritable effect of these two factors.

Crosses between certain Drosophila melanogaster strains may give rise to female sterility of non-Mendelian determination. Reduced fertility is observed in F1 females, known as SF females, from crosses between females of "reactive" strains and males of "inducer" strains. The extent of this reduction of fertility depends on the strains which are used in the cross and on two non-genetic factors: age and temperature. The fertility of SF females increases with ageing. Also, exposing them for a short period to a high temperature (29 degrees C) either increases or decreases the probability of hatching of the eggs according to the stage of oogenesis at which the heat treatment is applied. A very striking point is that qualitatively quite similar, though attenuated, effects are observed when the two factors (ageing and temperature) are applied not directly to SF females, but to their maternal ancestors: mothers and grandmothers.     

Non-Mendelian Female Sterility in DROSOPHILA MELANOGASTER: Influence of Aging and Thermic Treatments. III. Cumulative Effects Induced by These Factors

Crosses between various strains of Drosophila melanogaster may give rise to a female sterility of non-Mendelian determination. Reduced fertility is observed in females, known as SF females, bred from crosses between females of "reactive" strains and males of "inducer" strains. The reduced fertility of the SF females is the result of an interaction between an extrachromosomal property varies considerably in its ability to reduce fertility. The fertility reduction of the SF females corresponds to what is known as the reactivity level of their reactive mothers. Two nongenetic factors can modify the level of reactivity: aging and temperature. The action of aging is cumulative. When the flies of a reactive strain are submitted at each generation to the action of this factor, the level of reactivity of this strain is gradually modified. The modifications induced are reversible. Indeed, when such a modified strain is returned to standard breeding conditions, the reactivity returns progressively to its initial level. The effect of thermic treatments also seems to be cumulative and reversible.     

Specific expression of the Drosophila midline-jumper retro-transposon in embryonic CNS midline cells

Here we describe of a novel Drosophila LTR-type retrotransposon that is expressed in the embryonic CNS midline glia and in the embryonic germ cells. The element is related to the gypsy and burdock retrotransposons and was termed midline-jumper. In addition to cDNA clones generated from internal retrotransposon sequences, we have identified one cDNA clone that appears to reflect a transposition event, indicating that the midline-jumper retrotransposon is not only transcribed but also able to transpose during Drosophila development.