A Mutational Analysis of the Triplo-Lethal Region of DROSOPHILA MELANOGASTER

The extensive analysis of the impact of segmental aneuploidy by Lindsley et al. (1972) showed that there are relatively few haplo-lethal loci in the genome and that, with one exception, all loci are triplo-viable. The exceptional locus, which lies in salivary gland chromosome region 83D-E, is associated with lethality when present in either one or three doses in an otherwise diploid individual (Denell 1976). The genetic nature of the phenomenon has been studied by examining the rates of induction, by ionizing radiation and chemical mutagens, of mutations affecting the dose-sensitive behavior. For both types of mutagens, the frequency of inactivation of the locus is relatively low, and a high proportion of such mutations is associated with chromosomal deficiencies. These data indicate that the locus is infrequently and perhaps never inactivated by a DNA base-pair substitution and thus that the triplo-lethal phenomenon is not associated with a "typical" structural gene. It is possible that the triplo-lethal locus is very small, is reiterated or otherwise complex or is functionally insensitive to base-pair substitutions. The result that all mutations that complement a duplication of the triplo-lethal locus are lethal in heterozygous combination with a normal third chromosome argues that triplo- and haplo-lethality are concomitants of the same phenomenon. Salivary gland chromosome analysis of newly induced deficiencies and duplications localizes the locus to 83D4,5--83E1,2, and further cytogenetic mapipulation shows that the dose-sensitive behavior is independent of the position of the locus in the genome.     

Analysis of the Cut Locus of DROSOPHILA MELANOGASTER

Mutants of the cut (ct) locus can be divided into two classes: viable and lethal. Most of the viable alleles are characterized by varying degrees of scalloping and notching of the wings. One mutant, kinked femur, exhibits kinking of the femurs and failure of wing expansion, but no other changes in wing structure. In heterozygous combination with the other viable alleles, it exhibits complete complementation, but it fails to complement with lethal ct alleles with respect to its viable phenotype. Similarly, all of the other viable ct alleles express a mutant wing phenotype when heterozygous with lethal ct alleles.-Mapping experiments indicate that the lethal alleles, which comprise the majority of all ct mutations recovered, are confined to a small region at the right end of the locus. That this restriction is real and not an artifact imposed by the limited number of lethal mutations mapped in the locus is supported by an examination of the mutant ct(JC20), a presumptive deficiency for the left-most third of the locus. Despite its behavior as a deletion, ct(JC20) is viable, though mutant, in combination with the lethal alleles. The restriction of the noncomplementary lethals to a small part of the locus, distinct from the other ct mutants, suggests a polarity that may define a segment that functions only in cis within the complex.-Based on the comparison of the data with the prediction of several models, we suggest that the left portion of the locus, which contains the viable alleles, defines a regulatory region controlling the expression of the locus, while the segment encoding a polypeptide product is at the right end and only it is capable of mutating to a lethal state.     

Genetic Analysis of the Hairy Locus in DROSOPHILA MELANOGASTER

Mutations of the hairy locus in Drosophila may affect both adult chaeta differentiation and embryonic segmentation. In an effort to understand this phenotypic complexity, we have analyzed 30 mutant alleles of the locus. We find that the alleles fall into four groups according to their complementation properties, suggesting a structurally complex locus in which two distinct functions share a common coding region.     

Cytogenetic Analysis of the Chromosomal Region Immediately Adjacent to the Rosy Locus in DROSOPHILA MELANOGASTER

This report describes the genetic analysis of a region of the third chromosome of Drosophila melanogaster extending from 87D2–4 to 87E12–F1, an interval of 23 or 24 polytene chromosome bands. This region includes the rosy (ry, 3–52.0) locus, carrying the structural information for xanthine dehydrogenase (XDH). We have, in recent years, focused attention on the genetic regulation of the rosy locus and, therefore, wished to ascertain in detail the immediate genetic environment of this locus. Specifically, we question if rosy is a solitary genetic unit or part of a larger complex genetic unit encompassing adjacent genes. Our data also provide opportunity to examine further the relationship between euchromatic gene distribution and polytene chromosome structure.——The results of our genetic dissection of the rosy microregion substantiate the conclusion drawn earlier (Schalet, Kernaghan and Chovnick 1964) that the rosy locus is the only gene in this region concerned with XDH activity and that all adjacent genetic units are functionally, as well as spatially, distinct from the rosy gene. Within the rosy micro-region, we observed a close correspondence between the number of complementation groups (21) and the number of polytene chromosome bands (23 or 24). Consideration of this latter observation in conjunction with those of similar studies of other chhromosomal regions supports the hypothesis that each polytene chromosome band corresponds to a single genetic unit.     

A Genetic and Cytogenetic Analysis of the Region Surrounding the Lsp-1 beta-Gene in DROSOPHILA MELANOGASTER

The region surrounding the gene coding for the β-polypeptide (21D-22C) of the major Drosophila melanogaster larval serum protein, LSP-1, has been studied in detail. Seven new γ-ray-induced deficiencies of the region have been used, together with the two extant deficiencies, to map the position of the β-gene and of the 55 newly induced ethyl methanesulfonate mutants uncovered by one of the largest deficiencies. No lethal mutation of the β-gene was found.     

Evidence for a Sex-Linked Haplo-Inviable Locus in the Cut-Singed Region of DROSOPHILA MELANOGASTER

Many cytologically normal and rearranged cut mutants have been reported, but no known deficiency involves both ct and its close neighbor, singed. This fact prompted an investigation of the mutational response of the ct–sn interval. Approximately 24,000 F₁ female progeny of 7-day-old males given 2000 or 3000r X-ray exposures were examined for the presence of newly induced mutations at the Notch, carmine, ct, and sn loci. One sn, 2 cm-ct, 31 N, and 33 ct mutants were found, indicating that the frequency of recovery of ct mutants is much greater than that of either cm or sn, as high even as N. Among the F₁ female progeny were two deficiency mutants that expressed both cm and ct (separated by 21 bands), but none expressed both ct and sn (separated by only 14 bands). Of the 18 cytologically analyzed ct mutants, two proved to be deficiencies; neither extended farther to the right than 7C1. No reported ct deficiency extends with certainty farther to the right than 7C4. This fact, together with the scarcity of sn deficiencies, suggests the presence of a haplo-insufficient locus between ct and sn that prevents recovery of ct-sn deficiencies. The analysis of the deficiency component of Tp sn^S93, a short transposition which moves most of the ct–sn interval from 7BC to 8D, proved the existence, just to the left of sn, of a haplo-inviable locus that prevents the development of females heterozygous for its deficiency.—A marked similarity between mutants at the N and ct loci was noted.     

Genetic and Developmental Analysis of the Locus vnd in DROSOPHILA MELANOGASTER

Genetic and developmental analysis of an X-linked vital locus vnd was undertaken. Embryos hemizygous for the original allele vnd did not hatch and exhibited a disorganized ventral nervous system (VNS). The mutation maps in the region 1B6-7 to 1B9-10, a subregion of an area previously shown to be essential to normal neural development. In this paper, we report isolation of five new alleles at the locus vnd. Genetic complementation analysis of all mutations at the vnd locus, with lethal alleles at adjacent loci, indicates that all lesions at the locus vnd affect only one vital gene function in the region. Four of the five alleles are embryonic lethal; one allele is subvital and behaves like an hypomorphic mutation. Hemizygous embryos for three of the four embryonic lethal alleles were inspected in histological sections; all exhibited disorganized VNS similar to the original allele. The developmental analysis in gynandromorphic genetic mosaics shows that (1) vnd+ gene function is not essential in most imaginal-disc cell derivatives, (2) only about 30% of the mosaic zygotes survive as adults, (3) mosaic zygotes with mutant tissue close to the head cuticle are least likely to survive, and (4) mutant tissue in the thoracic ganglion in the adult is not necessarily lethal. The mosaic data are consistent with the vnd+ gene function being necessary in neural cells derived from the anterioventral region of the blastoderm.     

Negative Complementation at the Notch Locus of DROSOPHILA MELANOGASTER

Four Abruptex alleles (Ax^E1, Ax^E₂, Ax^₉B₂, and Ax¹⁶¹⁷²) have been mapped within the Notch locus. Based on their visible phenotypes and their interactions with one another and with N mutations, the Ax alleles can be divided into two groups. Heterozygous combinations of members of the same group are intermediate in phenotype compared to the respective homozygotes, whereas heterozygotes of Ax alleles from different groups exhibit negative heterosis, being much less viable and more extremely mutant than either homozygote. It is suggested that the Notch locus is a multi-functional regulator ("integrator") gene, whose product possesses both "repressor" and "activator" functions for the processes it regulates.     

Temperature-Sensitive Mutations of the Notch Locus in DROSOPHILA MELANOGASTER

Temperature-conditional mutations of the Notch locus were characterized in an attempt to understand the organization of a "complex locus" and the control of its function in development. Among 21 newly induced Notch alleles, about one-half are temperature-conditional for some effects, and three are temperature-sensitive for viability. One temperature-sensitive lethal, l(1)N^ts1, is functionally non-complementing for all known effects of Notch locus mutations and maps at a single site within the locus. Among the existing alleles involved in complex patterns of interallelic complementation, Ax^59d5 is found to be temperature-sensitive, while fa ^g, spl, and l(1)N are temperature-independent. Whereas temperature-sensitive alleles map predominantly to the right-most fifth of the locus, fa^g, spl, and l(1)N are known to map to the left of this region. Temperature-shift experiments demonstrate that fa^g, spl, and l(1)N cause defects at specific, non-overlapping times in development.—We conclude (1) that the Notch locus is a single cistron (responsible for a single functional molecule, presumably a polypeptide); (2) that the right-most fifth of the locus is, at least in part, the region involved in coding for the Notch product; (3) that the complexity of interallelic complementation is a developmental effect of mutations that cause defects at selected times and spaces, and that complementation occurs because the mutant defects are temporally and spatially non-overlapping; and (4) that mutants express selected defects due to critical temporal and spatial differences in the chemical conditions controlling the synthesis or function of the Notch product. The complexity of the locus appears to reside in controlling the expression (synthesis or function) of the Notch product in development.     

Molecular Mapping of the ROSY Locus in DROSOPHILA MELANOGASTER

The DNA from the chromosomal region of the Drosophila rosy locus has been examined in 83 rosy mutant strains. Several spontaneous and radiation-induced alleles were associated with insertions and deletions, respectively. The lesions are clustered in a 4-kb region. Some of the alleles identified on the DNA map have been located on the genetic map by fine-structure recombination experiments. The genetic and molecular maps are collinear, and the alignment identifies the DNA location of the rosy control region. A rosy RNA of 4.5 kb has been identified; its 5' end lies in or near the control region.     

Frequency-Dependent Selection at the LAP Locus in DROSOPHILA MELANOGASTER

Results of fitness estimates for the Lap locus in Drosophila melanogaster revealed that under crowded media conditions gene frequency equilibrium was maintained by frequency-dependent selection. Evidence was obtained that indicated that mating and egg-to-adult viability were frequency dependent.     

Excess Polymorphism at the Adh Locus in DROSOPHILA MELANOGASTER

The evolutionary history of a region of DNA encompassing the Adh locus is studied by comparing patterns of variation in Drosophila melanogaster and its sibling species, D. simulans. An unexpectedly high level of silent polymorphism in the Adh coding region relative to the 5' and 3' flanking regions in D. melanogaster is revealed by a populational survey of restriction polymorphism using a four-cutter filter hybridization technique as well as by direct sequence comparisons. In both of these studies, a region of the Adh gene encompassing the three coding exons exhibits a frequency of polymorphism equal to that of a 4-kb 5' flanking region. In contrast, an interspecific sequence comparison shows a two-fold higher level of divergence in the 5' flanking sequence compared to the structural locus. Analysis of the patterns of variation suggest an excess of polymorphism within the D. melanogaster Adh locus, rather than lack of polymorphism in the 5' flanking region. An approach is outlined for testing neutral theory predictions about patterns of variation within and between species. This approach indicates that the observed patterns of variation are incompatible with an infinite site neutral model.     

Position Effects at the Hairy Locus in DROSOPHILA MELANOGASTER

Radiation-induced chromosomal rearrangements of h(+) have given rise to several Drosophila stocks that exhibit apparent position-effect inactivation; i.e., flies carrying the rearranged chromosomes heterozygously with h show varying degrees of hairiness. The numbers of hairy chaetae produce a quantifiable index of position effect. Six such "position-allele" stocks are here discussed, both as to their basic expressions and in all possible pair-wise combinations with each other. Such crosses reveal complex interactions between the respective position alleles; little evidence is seen for clear-cut dominance or recessiveness. The stocks appear not to conform unequivocally to classical distinctions between variegated and stable types of position effects, nor to usual dicta relating the degree of inactivity to the proximity to heterochromatin. Indeed, these stocks appear to suggest additional dimensions to several of the principles to which position effects usually subscribe. The evidence additionally suggests that the hairy locus itself is associated with a tissue-specific suppressor effect on an otherwise polygenic system that produces the chaetae associated with the hairy phenotype.     

Allelic Negative Complementation at the Abruptex Locus of DROSOPHILA MELANOGASTER

The mutations of the Abruptex locus in Drosophila melanogaster fall into three categories. There are recessive lethal alleles and viable alleles. The latter can be divided into suppressors and nonsuppressors of Notch mutations. The recessive lethals are lethal in heterozygous combination with Notch. As a rule the recessive lethals are lethal also in heterozygous combination with the viable alleles. Heterozygous combinations of certain viable alleles are also lethal. In such heterozygotes, one heteroallele is a suppressor of Notch and the other is a nonsuppressor. Other heterozygous combinations of viable alleles are viable and have an Abruptex phenotype. The insertion of the wild allele of the Abruptex locus as an extra dose (carried by a duplication) into the chromosomal complement of the fly fully restores the viability of the otherwise lethal heterozygotes if two viable alleles are involved. The extra wild allele also restores the viability of heterozygotes in which a lethal and a suppressor allele are present. If, however, a lethal and a nonsuppressor are involved, the wild allele only partly restores the viability, and the effect of the wild allele is weakest if two lethal alleles are involved. It seems likely that of the viable alleles the suppressors of Notch are hypermorphic and the nonsuppressors are hypomorphic. The lethal alleles share properties of both types, and are possibly antimorphic mutations. It is suggested that the locus is responsible for a single function which, however, consists of two components. The hypermorphic mutations are defects of the one component and the hypomorphic mutations of the other. In heterozygotes their cumulative action leads to decreased viability. The lethal alleles are supposed to be defects of the function as a whole. The function controlled by the locus might be a regulative function.     

Mutational Events in the Triplo- and Haplo-Lethal Region (83de) of the DROSOPHILA MELANOGASTER Genome

The Drosophila melanogaster genome contains a single region (at 83DE on the polytene chromosome map) for which both heterozygous deficiency and heterozygous duplication are inviable. Seven EMS-induced mutations have been recovered that are viable in combination with a duplication of this region. Two classes of mutations are reported: (1) Mutations that allow survival of flies with either a duplication or a normal third chromosome. These mutations retain Ki, a closely linked marker on the mutagenized chromosome. They fail to complement, and one has been mapped to the vicinity of 83DE. (2) Mutations that allow survival only in heterozygous combination with a duplication and have lost the Ki marker. These mutations represent new deletions of the dose-sensitive information. The possible structural organization of the 83DE region is discussed in light of these two classes of mutations.