A mutable white-inversion in Drosophila melanogaster

Summary From a zeste mutant stock with a mutable white locus a new mutant (z w ^w ) was isolated. It has a white-eyed phenotype and a short X-chromosome inversion (In(1)w ^w ) which extends from salivary chromosome bands 3B2-C1 to 4B4-C1. In giant chromosomes of heterozygotes the inversion is unusually tightly paired. Probably because of this intimate pairing the recombination frequencies for regions near the inversion are not decreased in comparison to those for structurally normal chromosomes. The inversion chromosome is mutable. The mutations which arise have pigmented eyes and can be subdivided into two groups. One group is characterized by a re-inversion to normal chromosome structure. The mutability of the white locus appears to be independent of the inversion and reinversion. The process of reinversion is discussed.     

Maternal influence upon the V-type gene position effect in Drosophila melanogaster.

Summary We have studied the influence of various factors on the V-type position effect of the white gene transposed to heterochromatin as a result of different chromosome rearraugements in D. melanogaster. Variegation due to the white gene position effect is much weaker if the flies have received Dp(1;3)w^vco from parental males, and not females. The origin of the chromosome rearrangement does not have this influence in the case of T(1;4)w^m5 or has it to insignificant extent in the case of In(1)w^m4. The Y-chromosome in maternal genome strongly suppresses Dp(1;3)w^vco-induced variegation even in the progeny which has not received an extra Y-chromosome but only if this progeny gets Dp(1;3)w^vco from the same female. The low temperature (16° C) at which parental females have developed, considerably affects the position effect in the progeny with Dp(1;3)w^vco, whereas the temperature of males' development has no influence at all. The maternal temperature effect occurs also when Dp(1;3)w^vco has come down from the father, though it is stronger if the mother subjected to low temperature treatment bore the rearrangement. The influence of temperature seems to take effect at the final stages of oogenesis.The data obtained lead one to suppose that the influence of genotypic and external factors on variegation is passed to the next generation of flies in different ways. The direction of crosses and additional Y-chromosome heterochromatin in the maternal genome seem to affect variegation in the progeny through changes in the properties (structure) of the chromosome rearrangement expressing the position effect. As to the temperature of the mothers development, only a small part of its influence may be accounted for by the same mechanism, whereas most of the temperature influence seems to be passed on through other components of the nucleus or through the cytoplasm.     

An analysis of white-mottled mutants inDrosophila hydei,with observations on X-Y exchanges in the male

Chromosomes and phenotypes of four different sex-linkedwhite-mottled mutants of the position-effect variogation type were studied. Three mutants (w ^m1,w ^m2,w ^m3) are X-chromosomal rearrangements which shift the w⁺ locus into a position close to heterochromatin, but which have different ouchromatic and heterochromatic breaks. The fourth, a spontaneous derivative ofw ^m1, is an insertional duplication of part of the X chromosome, including thew ⁺ andN ⁺loci. The duplicated segment is inserted into the distal part of the long arm of the heterochromatic Y chromosome. It is designated,w ^m CoY, orXw ^m Co when transferred to the X chromosome.Three chromosomal types (w ^m1,w ^m CoY) and (Xw ^m Co) having the same cuchromatic break near thew ⁺ locus, cause large-spotted eyes whereas two others (w ^m2,w ^m3) produce a popper-and-salt type of mottling. From the position of the various eu- and heterochromatic breaks, it appears that the distance of thew ⁺ locus to the point of reunion with heterochromatin, rather than the amount or type of adjoining heterochromatin, dietates the phenotypic action of the displacedw ⁺ locus, in the sense of a spreading effect on two proposed functional subunits within thew ⁺ locus.The pigmentation background against which the mottling effect is produced, i.e., a givenw-allele with its characteristic colour, or other eye colour mutations, does not seem to affect the type of mottling. Drosopterins and ommochromes react in the same way to modifing factors like temperature and supernumerary Y chromosomes. Two mutants (w ^m2 andw ^m CoY) while reacting in the same manner to Y chromosomes showed an opposite temperature response.By exchange between the heterochromatin of the Y and X chromosome inw/w ^m CoY males thew ^m Co duplication was transferred between the sex chromosomes with a certain regularity. It is not yet known wether the exchanges are mitotic or meiotic in origin but their heterochromatic nature has been demonstrated cytologically.     

FB elements can promote exon shuffling: a promoter-less white allele can be reactivated by FB mediated transposition in Drosophila melanogaster

Foldback (FB) elements are transposable elements found in many eukaryotic genomes; they are thought to contribute significantly to genome plasticity. In Drosophila melanogaster, FBs have been shown to be involved in the transposition of large chromosomal regions and in the genetic instability of some alleles of the white gene. In this report we show that FB mediated transposition of w ^67C23, a mutation that deletes the promoter of the white gene and its first exon, containing the start codon, can restore expression of the white gene. We have characterized three independent events in which a 14-kb fragment from the w ^67C23 locus was transposed into an intron region in three different genes. In each case a local promoter drives the expression of white, producing a chimeric mRNA. These findings suggest that, on an evolutionary timescale, FB elements may contribute to the creation of new genes via exon shuffling.Electronic Supplementary Material Supplementary material is available in the online version of this article at Communicated by G. P. Georgiev     

The action of the Notch locus in Drosophila hydei

The phenotypic effects of different doses of the dominant, sex-linked mutant Notch (N) and its wildtype allele (N ⁺) were studied in Drosophila hydei, N being lethal in homozygous or hemizygous condition. Various dosage combinations were made by using N ⁺ N and N ⁺ N ⁺ attached-X chromosomes as well as X and Y N ⁺-duplication chromosomes (w ^mCoY, Xw^mCo,and DpCo ^Nt). The N mutant used, N ⁶⁸, is associated with a small inversion: In (I) N ⁶⁸.The wing phenotype was found to depend solely on the number of functional (N ⁺) alleles present, irrespective of the dose of N. Females with a single dose of N ⁺ are phenotypically Notch, females with three or four doses of N ⁺ show a Confluens wing phenotype. The latter occurs in varying degrees of expression which seem to be correlated with the relative amounts of sex-chromosomal heterochromatin present. In males the N ⁺ locus behaves as a dosage compensated locus either on the X or the Y chromosome.In the w ^mCo (w⁺N⁺) duplication, the w ⁺ locus shows variegation when placed over white, whereas N ⁺ placed over N ⁶⁸ does not. The former being situated closer to the heterochromatin in this aberration, this is consistent with the idea of gene inhibition by heterochromatin but at the same time would imply a very limited spreading effect.     

X-Y exchanges in males of Drosophila hydei carrying the w m Co duplication

Males carrying, inserted on their Y chromosome, a small fragment of X including the w ⁺ (and N ⁺) locus (white-mottled Confluens, w ^m Co), were crossed with the purpose of scoring exceptional progeny. Some of the male and female exceptions were progeny tested and further analysed. Among the various mechanisms which may lead to exceptional offspring, X-Y exchanges proved to occur with a not negligible frequency. The rate was 3%. Nondisjunction accounts for the bulk of the remaining exceptions and appears to be increased considerably in the presence of rearrangements on one or the other of the sex chromosomes.The w ^m Co fragment after having been switched from Y to X by some mechanism other than regular crossing over, may become retransferred to a normal Y chromosome, but at a rate below 3%.     

Genetic instability in Drosophila melanogaster. Evidence for insertion mutations

Summary An X chromosome in Drosophila melanogaster is described which is mutationally unstable. Mutational events were identified through phenotypic changes associated with a tandem duplication of the X chromosome in which the white locus is present in duplicate. The left segment of the tandem duplication was marked with the mutant w ^sp, the right segment with mutant w ^17G. Some of the phenotypic changes were identified as deletions involving the w ^17G marked segment of the duplication. Other phenotypic changes involved the left segment in which phenotypically w ^sp mutated to w. Experimental evidence is presented which attributes these latter mutations to insertions of foreign DNA into the w locus equivalent to the insertion mutations of E. coli.     

The genetic basis of resistance and sensitivity to the meiotic drive gene D in the mosquito Aedes aegypti L.

A study has been made on the genetic basis of meiotic drive at the Distorter (D) locus which, in coupling with the male-determining gene (or region) M on the Y chromosome, causes production of excess male progeny. Its effect is regulated by the sensitivity/resistance of the X chromosome. This study demonstrates that there are two major loci controlling resistance/sensitivity to MD: (1) the m gene (or region) on the X chromosome (allelic with M) which may be either m ^R or m ^S (resistant or sensitive), (2) the t (tolerance) gene (or genes) which recombines with m and, if present, largely counteracts the effect of m ^S . There is also evidence that MD itself is capable of limited adaptation.The conclusions were derived from using MD males of the T30 or ACCRA strains (from Trinidad and Ghana respectively). The work involved the use of the CHIPEI and RED strains with sensitive X chromosomes, the latter also carrying the t (tolerance) gene which is linked to re (red eye) and m (the sex-determining locus or region) but recombines with both. The implications of these findings for using MD as a method of population control are discussed.     

Y chromosome-specific mutations induced by a giant transposon in Drosophila hydei

Summary The w ^m Co duplication of Drosophila hydei (Dp (1; Y) 16B2-17B1) contains 13–16 bands in salivary gland chromosomes. The duplication resides preferentially in the X heterochromatin or on the Y chromosome. In some stocks frequent (up to 4×10^-3) exchanges of the duplication occur between different Y chromosomes (T(X; Y) and free Y) or between the X and the Y chromosome. About 60% of the T(X; Y)-Y exchanges induce mutations in the Y chromosomal male fertility genes of the recipient Y chromosome. From the mutational spectrum generated by the T(X; Y)-Y transpositions and from the variable efficiency as acceptor of different X-Y translocations it can be concluded that the exchanges show a remarkable site specificity: distal positions in the long arm of the Y chromosome are occupied preferentially. More proximal positions in the long arm of insertions into the short arm of the Y chromosome are found only with a lower frequency. No transpositions to the autosomes have been recovered. Duplications are lost with highly differing frequencies. The losses are not linked with insertions of the w ^m Co element into a new position and are more frequent than transpositions. Therefore, we regard the w ^m Co element as a giant transposon.     

The genetic analysis of a large transposing element of Drosophila melanogaster

A member of Ising's family of large transposing elements (TEs) has inserted into, or very near, the crinkled (ck, 2–50) locus. This TE (TE36) carries functional alleles of both the white and roughest loci, and causes a hypomorphic mutation of ck. The TE is visible in polytene chromosomes as a two-banded insertion between 35B9 and 35C1. These bands show homology to foldback (FB) elements by in situ hybridization. All spontaneous losses of TE36 remain mutant for ck and retain sequences homologous to FB at the site of TE's insertion. TE36 carries only one functional copy of w ⁺, by the criterion that z w, TE36/ + flies are wild-type for eye color but z w; TE36/TE36 flies are zeste. This white⁺ gene is dosage compensated since w/Y; TE36/+ males have twice as much eye pigment as w/w; TE36/ + females. A form of the TE that has four polytene chromosome bands and expresses twice as much pigment as TE36 has been recovered. However, its white genes are not suppressed by zeste.     

H3S10 phosphorylation by the JIL-1 kinase regulates H3K9 dimethylation and gene expression at the white locus in Drosophila

The JIL-1 kinase is a multidomain protein that localizes specifically to euchromatin interband regions of polytene chromosomes and is the kinase responsible for histone H3S10 phosphorylation at interphase. Genetic interaction assays have suggested that the function of the epigenetic histone H3S10ph mark is to antagonize heterochromatization by participating in a dynamic balance between factors promoting repression and activation of gene expression as measured by position-effect variegation (PEV) assays. Interestingly, JIL-1 loss-of-function alleles can act either as an enhancer or indirectly as a suppressor of w^m4 PEV depending on the precise levels of JIL-1 kinase activity. In this study, we have explored the relationship between PEV and the relative levels of the H3S10ph and H3K9me2 marks at the white gene in both wild-type and w^m4 backgrounds by ChIP analysis. Our results indicate that H3K9me2 levels at the white gene directly correlate with its level of expression and that H3K9me2 levels in turn are regulated by H3S10 phosphorylation.     

Genes controlling chromosome activity

Normal propagation of Y chromosome lampbrush loops was used as a screening tool in order to recover X-linked mutations controling Y chromosome activation. The nature of the most extreme mutationthus recovered, sterile (1) XL2, is described. It is a recessive gene mutation, readily mapped 2 cross over units distally to white. The mutation exerts its sterilizing effect by blocking normal unfolding of all Y lampbrush loops, but does not affect the unique shape of each diminutive loop. The degree to which a loop forming site is developed is partially temperature sensitive. It is independent however, on its map location or the dose of homologous as well as heterologous sites. It was provisionally concluded therefore that site response to the XL2 effect is a stage specific and not a quantitative one. The possible ways by which non homologous genes control Y chromosome activity are discussed.     

Volatile general anesthetics reveal a neurobiological role for the white and brown genes of Drosophila melanogaster

Molecular and cellular evidence argues that a heterodimer between two ABC transporters, the White protein and the Brown protein, is responsible for pumping guanine into pigment‐synthesizing cells of the fruit fly, Drosophila melanogaster. Previous studies have not detected White or Brown outside pigment‐synthesizing cells nor have behavioral effects of null mutants been reported, other than those that are visually dependent. Nevertheless, we show here that exposure to the volatile general anesthetic (VGA) enflurane reveals a difference in neuromuscular performance between wild‐type flies and those that carry a null allele in either the white or brown gene. Specifically, in a test of climbing ability, w¹¹¹⁸ or bw¹ flies are much less affected by enflurane than are congenic controls. Altered anesthetic sensitivity is still observed when visual cues are reduced or eliminated, arguing that white and brown contribute to neural function outside the eye. This hypothesis is supported by the detection of white message in heads of flies that are genetically altered so as to lack pigment‐producing cells. The w¹¹¹⁸ or bw¹ mutations also alter the response to a second VGA, halothane, albeit somewhat differently. Under some conditions, the combination of w¹¹¹⁸ with another mutation that affects anesthesia leads to a drastically altered phenotype. We consider several ways by which diminished transport of guanine could influence neural function and anesthetic sensitivity. Published 2001 John Wiley & Sons, Inc. J Neurobiol 49: 339–349, 2001     

Molecular lesions associated with white gene mutations induced by I-R hybrid dysgenesis in Drosophila melanogaster

We have identified molecular lesions associated with six mutations, w^IR2 and w^IR4-8, of the white gene of Drosophila melanogaster. These mutations arose in flies subject to I-R hybrid dysgenesis. Four of the mutations give rise to coloured eyes and are associated with insertions of 5.4-kb elements indistinguishable from the I factor controlling I-R dysgenesis. The insertion associated with w^IR4 is at a site which, within the resolution of these experiments, is identical to that of two previously studied I factors. This appears to be a hot-spot for I factor insertion. We have compared the sites of these insertions with sequences complementary to white gene mRNA identified by Pirrotta and Bröckl. The hot-spot is in the fourth intron. The insertion carried by w^IR5 is either within, or just beyond, the last exon. The insertion carried by w^IR6 is near the junction of the first exon and first intron. The w^IR2 mutation is a derivative of w¹. It contains an insertion of I factor DNA within, or immediately adjacent to, the F-like element associated with w¹, and results in restoration of some eye colour. This insertion is just upstream of the start of the white mRNA. Mutations w^IR7 and w^IR8 are deletions removing mRNA coding sequences. Both determine a bleached white phenotype.     

Third chromosome suppressor of position-effect variegation loci in Drosophila melanogaster

Summary As a result of a genetic analysis of 63 third chromosome suppressor mutations of position-effect variegation 12 different loci showing dominant suppression have been identified and their map positions determined. A compilcation of the genetic data available for each suppressor locus is given. The strong suppressor effects of the mutations have been quantified by measurements of white variegation inw ^m4h /w ^m4h ,w ^m4h /Y andw ^m4h /O flies. Mutant alleles of three loci were found in these studies to dominate over the strong enhancer effect of complete loss of the Y chromosome. Most of the identified loci suppressing position-effect variegation represent essential genetic funtions; only three loci represent nonessential functions. Mutations of two loci display recessive butyrate sensitivity and lethal interaction with the heterochromatic Y chromosome suggesting that these genes affect chromosomal condensation. Studies with deficiencies and triploids revealed that most of the loci represent haplo-abnormal suppressor functions. The use of the isolated mutant material for genetic, developmental and molecular studies of processes connected with gene inactivation in position-effect variegation is discussed.Dedicated to Prof. H.J. Becker on the occasion of his 6th birthday     

Bestimmung des Heterochromatisierungsstadiums beim white-Positionseffekt mittels r?ntgeninduzierter mitotischer Rekombination in der Augenanlage von Drosophila melanogaster

Summary Position-effect variegation of eye pigmentation in the examined Dp(1;3)N ^264-58 females is due to an insertion of a X-chromosome section including the white-locus into the proximal heterochromatic region of the third chromosome. The light and dark pigmented areas have a cell lineage basis (Fig. 2). Flies bearing the w ⁺-duplication had two X-chromosomes marked with w ^a lz ⁵⁰ e and w ^a rb rux ² respectively (Fig. 3). X-ray induced mitotic recombination in presumptive eye cells of larvae resulted in w ^a lz ^50e /w ^a rb rux ² twin mosaic spots in the adult eyes. After young larvae were treated twin spots appeared, which had one partner light colored and one dark. Such combinations were rarely found after older larvae were treated. Treatment of young larvae in addition produced twin spots with one or both partners variegated (Figs. 5 and 6). Sometime after the stage at which younger larvae were treated and before the stage at which older larvae were treated the translocated w ⁺-gene in each cell was determined for function or no function. As a result the progeny of each of these cells synthesized pigment or not during the pupal stage. At a temperature of 25.5° C the developmental phase during which determination, i.e. heterochromatization of the white gene, takes place, begins not earlier than 39 hours after egg laying and ends about 8 hours later (Fig. 7). In females heterozygous for the short arm of the heterochromatic Y-chromosome linked distally to the X-chromosome (Y ^S X/X) one twin spot partners is homozygous for this arm (Y ^SX/Y^S X), the other lacks it (X/X; Fig.4a). The Y ^SX/Y^S X-partner were more frequently dark pigmented than the X/X-partners (Tables 3 and 4). This shows that heterochromatization of the translocated w ⁺-genes is markedly influenced by the genotype of the single cell. When two genotypes with varied amounts of heterochromatin were compared (Fig. 4) no difference in the phases of heterochromatization could be observed (Table 5). Therefore, when position-effect variegation is modified by varying the amount of heterochromatin in the genome the modification is probably not due to a shift in the phase of heterochromatization.

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Vorgelegt von E. Hadorn     

Influence of deficiency of the histone gene-containing 38B-40 region on X-chromosome template activity and the white gene position effect variegation in Drosophila melanogaster.

Summary The deficiency of the 38B-40 region containing histone genes in one of the 2nd chromosomes of D. melanogaster triploid intersexes increases the template activity of X-chromosomes both in vivo and in vitro without noticeably affecting autosome activity. This deficiency in the heterozygous state inhibits the variegated position effect of the white gene in the T(1;3)w ^vcotranslocation in diploid females and males, but not affect their rate of development. The variegation suppressor Su(var)hg-1 not only suppress the gene position effect in diploid flies, but also increases the template activity of X-chromosomes in triploid intersexes.The results are discussed with respect to the dependence of gene activity on the structure of chromosomes (density of DNP packing).