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.     

Analysis of the Negative Complementation of Abruptex Alleles in Gynandromorphs of DROSOPHILA MELANOGASTER

Certain Abruptex alleles in the Notch pseudoallelic series of Drosophila melanogaster show strong negative complementation. Heterozygous combinations of some viable alleles are lethal. As a lethal system this is unique. Analysis of this type of allelic interaction in gynandromorphs suggests that the lethal focus has a fate-map site in the anterior part of the fly, probably close to the central part of the thorax. In addition to the lethal effect, negative interaction of the alleles can also be seen in the morphogenesis of wings and chaetae of thorax and head. At this morphological level, the negative interaction of the alleles appears to be autonomous.     

Tissue-Specific and Complex Complementation Patterns in the Punch Locus of DROSOPHILA MELANOGASTER

Mutations in the Punch locus result in loss of GTP cyclohydrolase activity, but all mutations do not affect the enzyme in the same way. There are at least three classes of Punch mutations. One class results in a dominant eye color, recessive lethal phenotype. A second class of mutations also causes a recessive lethal phenotype, but heterozygous mutants have normal eye color. They show loss of GTP cyclohydrolase function in all tissues where activity can be measured. Alleles comprising a third class are recessive eye color mutations that are homozygous viable. Individuals with this third type of mutation show loss of enzyme activity in the eye, but show normal or near-normal activity elsewhere. In order to examine the organization and function of this locus further, we have performed interallelic complementation tests on 25 Punch mutations, monitoring viability and enzyme activity in prepupae and adults. Most allele combinations are lethal. Those that complement do so in ways that are tissue-or stage-specific and unpredictable. Tests of mutants with tissue-specific phenotypes and of individuals mutant for complementing Punch lethal alleles lead us to conclude that Punch is a complex locus, both with respect to its organization and to its products.     

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.     

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.     

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.     

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 Dumpy Complex Locus in DROSOPHILA MELANOGASTER: Complementation, Fine Structure and Function

An extensive genetic analysis of the dumpy locus is presented. This study includes complementation, fine structure mapping and allelic interaction. A number of complementing recessive lethals of the dp complex have been genetically mapped. Two alleles of the ol(v) type that complement l alleles map to the left portion of the locus. A number of olv alleles that complement both l and lv lethals map within the right portion of the locus.——Fine-structure analysis demonstrated that both olv and o alleles are distributed among various subloci. Evidence for spacer regions between subloci is presented.——An extensive discussion of the data considers whether the locus is unicistronic or multicistronic. The conclusion reached is that the locus is not a single functional cistron. The possibility of a single cistron encoding a multifunctional polypeptide is discussed.——The hypothesis is proposed that the left portion of the map and the l mutations function as regulatory sequences and that the right portion of the map encodes structural sequences.     

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.     

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.