Evidence for a Set of Closely Linked Autosomal Genes That Interact with Sex-Chromosome Heterochromatin in DROSOPHILA MELANOGASTER

It is proposed that there exists a special region in the euchromatin of the left arm of chromosome 2 (contained within sections 31-32 of the standard salivary gland chromosome map) that is defined by a set of genes, each one of which interacts with a specific sex-chromosome heterochromatic segment. The evidence for the existence of this region is, first, the exhibition, mapping, and analysis of five different maternal-effect, embryonic semi-lethals located in region 31-32. Secondly, in each case the consequence of the maternal effect is markedly influenced by the amount of X- or Y-chromosome heterochromatin carried by the progeny of mutant mothers. The nature of this interaction and possible reasons for the existence of the cluster of autosomal genes are discussed.     

Differential Response to Selection on the Two Sexes in DROSOPHILA MELANOGASTER

Artificial selection for short wing was performed in two Drosophila melanogaster populations with partially different gene pools: the C populations were derived from a Canton stock while the H lines were derived from a cross between Canton and a b, cn, vg strain. It is shown that in both populations selection on females (CF, HF) was more effective than selection on males (CM, HM). This difference cannot be explained in terms of differences in additive genetic variability between the two sexes because: (1) both sexes contribute to the genetic variability utilized by selection applied to one sex only, and (2) switching selection pressure on females in the M lines does not result in a response comparable to that obtained in the F populations; this rules out almost completely recombination as the responsible agent for the differences between the selection limits reached by M and F selections.-These results, together with several additional observations concerning sexual dimorphism, fitness and the effect of natural selection, suggest that a complex interaction should be involved in the differential response of M and F lines, controlling the wing length phenotype.     

Maternal-Zygotic Lethal Interactions in DROSOPHILA MELANOGASTER : Zeste-White Region Single-Cistron Mutations

Thirty-eight mutations in 13 essential loci in the zeste-white region were tested for interacting maternal and zygotic gene activity. Maternal mutant heterozygosity provided a partial maternal defect and position-effect variegation was used to alter the level of zygotic gene activity. This method yields a minimum estimate of the number of genes for which zygotic development depends upon both gene products stored in the egg and gene products synthesized in the zygote. Lethal interactions were found for one or more alleles at 10 of the 13 loci. The implications of these observations with respect to gene regulation and developmental sequence are considered.     

Spontaneous and Ethyl Methanesulfonate-Induced Mutations Controlling Viability in DROSOPHILA MELANOGASTER. I. Recessive Lethal Mutations

The efficiency of the adult feeding method for EMS treatment in Drosophila melanogaster was studied by measuring the frequency of induced recessive lethals on the second chromosome. The treatment was most effective when mature spermatozoa or spermatids were treated and was much less effective on earlier stages. The number of mutations induced was proportional to the concentration except at the highest doses. The recessive lethal rate was estimated to be about 0.012 per second chromosome per 10(-4) M. In addition, about 0.004-0.005 recessive lethals per 10(-4) M were found in a later generation in chromosomes that had not shown the lethal effect in the previous generation. When the experiments are done in a consistent manner and gametes treated as mature sperm or spermatids are sampled, the results are highly reproducible. However, modifications of the procedure, such as starvation before EMS treatment, can considerably alter the effectiveness of the mutagen.     

OM Mutations in DROSOPHILA ANANASSAE Are Linked to Insertions of a Transposable Element

It has been hypothesized that Om mutability in Drosophila ananassae (involving spontaneous mutation at 20 loci, resulting in semidominant, nonpleiotropic eye morphology defects) was due to insertion of a transposable element, tom. One particularly unstable X-linked Om allele produced several derivatives, one of which has a more extreme Om phenotype and was accompanied by a singed bristle mutant, sn ^9g. DNA probes from the sn locus of D. melanogaster were used to clone the homologous region of D. ananassae. Analysis of sn^9g DNA detected a 6.5-kb insert. Genomic Southern blotting and in situ hybridization techniques showed that this insert is repetitive and dispersed. The existence of the tom element is supported by genetic mapping that established homology between the 6.5-kb sn^9g insert and Om mutants at the four X-linked loci tested.     

Lethality Patterns and Morphology of Selected Lethal and Semi-Lethal Mutations in the Zeste-White Region of DROSOPHILA MELANOGASTER

Aspects of the developmental genetics of lethal and semi-lethal mutants representing 13 complementation groups (cistrons) in the 3A-3C region of the X chromosome of Drosophila melanogaster are given. Each of these cistrons is associated with a particular chromomere in the salivary gland chromosome. Mutants within each cistron have similar lethality patterns and morphological attributes, and the characteristics of a given cistron are distinct with respect to other cistrons. These results provide additional evidence that only one function is associated with each chromomere.-The results of the lethality pattern analysis are also compared with previous studies of lethal mutants of Drosophila.     

A Note on the Maternal Effect Mutants Daughterless and Abnormal Oocyte in DROSOPHILA MELANOGASTER

Two deficiencies for, and a dominant enhancer of, the second chromosome maternal effect mutant, "daughterless" (da), were induced with X-irradiation. Their properties were studied with respect to both da and the linked maternal effect mutant, "abnormal oocyte" (abo), with the following conclusions. (1) The most probable map positions of da and abo are: J-(1/2)-da-2(1/2)-abo, where J is a dominant marker located at 41 on the standard map. (2) The da locus is in bands 31CD-F on the polytene chromosome map; abo is to the right of 32A. (3) Because homozygous da individuals survive while individuals carrying da and a deficiency for da are lethal, it is concluded that da is hypomorphic. (4) From a weak da-like maternal effect in heterozygous da females induced by an "Enhancer of da," we have confirmed a previous report that (a) the amount of sex chromosome heterochromatin contributed by the father can influence the severity of the da maternal effect, and (b) the sex chromosome heterochromatin which influences the da effect is different from that which influences the abo effect. (5) The possibility that da and abo are in a special region of chromosome 2 concerned with the regulation of sex chromosome heterochromatin is strengthened by the observation that the Enhancer of da appears to rescue abnormal eggs produced by homozygous abo mothers. (6) The Enhancer of da is a translocation between chromosomes 2 and 3 with the second chromosome breakpoint in the basal heterochromatin; because the enhancing effect maps in this region of chromosome 2, it is possible that autosomal, as well as sex chromosomal, heterochromatin interacts with da and abo.     

Analysis of Y-Linked Mutations to Male Sterility in DROSOPHILA MELANOGASTER

Mating type in haploid cells of the yeast Saccharomyces cerevisiae is determined by a pair of alleles MATa and MAT alpha. Under various conditions haploid mating types can be interconverted. It has been proposed that transpositions of silent cassettes of mating-type information from HML OR HMR to MAT are the source of mating type conversions. A mutation described in this work, designated AON1, has the following properties. (1) MAT alpha cells carring AON1 are defective in mating. (2) AON1 allows MAT alpha/MAT alpha but not MATa/MATa diploids to sporulate; thus, AON1 mimics the MATa requirement for sporulation. (3) mata-1 cells that carry AON1 are MATa phenocopies, i.e., MAT alpha/mata-1 AON1 diploids behave as standard MAT alpha/MATa cells; therefore, AON1 suppresses the defect of mata-1. (4) AON1 maps at or near HMRa. (5) Same-site revertants from AON1 lose the ability to convert mating type to MATa, indicating that reversion is associated with the loss of a functional HMRa locus. In addition, AON1 is a dominant mutation. We conclude that AON1 is a regulatory mutation, probably cis-acting, that leads to the constitutive expression of silent a mating-type information located at HMRa.     

Interactions between FUSED and ENGRAILED, Two Mutations Affecting Pattern Formation in DROSOPHILA MELANOGASTER

In this paper we demonstrate that the severity of the engrailed phenotype is greatly increased when engrailed is combined with the X-linked mutation fused. The secondary sex combs of fu;en flies contain from two to four times more setae than do those of en siblings. The number of transverse rows of bristles on the metatarsus of the metathoracic leg is reduced by a factor of one and a half to two in fu;en flies when compared to their en siblings, while the frequency of reversed bract polarity, bristle abnormalities and misshapen metatarsi increases greatly. In addition, fu;en flies express the en wing phenotype more completely than their siblings and have a much higher frequency of wing vein abnormalities—some of which we have interpreted as triplications of the third and first longitudinal wing veins. We briefly discuss the significance of the fused-engrailed gene interaction.     

Induced Dominant Lethal Mutations and Cytotoxic Effects in Germ Cells of DROSOPHILA MELANOGASTER with Trenimon, Pdmt and Sodium Monofluorophosphate

Male and female Drosophila melanogaster with special sex chromosome or special autosome constitutions were fed with the mutagenic chemicals Trenimon (2,3,5-trisethyleneimino-1,4-benzoquinone) and PDMT (1-phenyl-3, 3-dimethyltriazene) and with the toxic substance Na2PO3F (sodium monofluorophosphate). The frequency of dominant lethality was recorded among the progeny. The results clearly show that dominant lethality is dose dependent for Trenimon- or PDMT-treated chromosomes in mature sperm and mature oocytes, and an increased amount of chromosomal material per nucleus yields an enhanced lethality. In contrast, a pure toxic effect of Na2PO3F on mature oocytes was demonstrated with one type of female. --With the stocks of Drosophila used, it is possible to distinguish between mutagenic and toxic effects of chemicals on the germ cells. Therefore, dominant lethality can be used as a simple and quick screening test for chemical mutagens.     

Multiple Allele Approach to the Study of Genes in DROSOPHILA MELANOGASTER That Are Involved in Imaginal Disc Development

The phenotypes of five different lethal mutants of Drosophila melanogaster that have small imaginal discs were analyzed in detail. From these results, we inferred whether or not the observed imaginal disc phenotype resulted exclusively from a primary imaginal disc defect in each mutant. To examine the validity of these inferences, we employed a multiple-allele method. Lethal alleles of the five third-chromosome mutations were identified by screening EMS-treated chromosomes for those which fail to complement with a chromosome containing all five reference mutations. Twenty-four mutants were isolated from 13,197 treated chromosomes. Each of the 24 was then tested for complementation with each of the five reference mutants. There was no significant difference in the mutation frequencies at these five loci. The stage of lethality and the imaginal disc morphology of each mutant allele were compared to those of its reference allele in order to examine the range of defects to be found among lethal alleles of each locus. In addition, hybrids of the alleles were examined for intracistronic complementation. For two of the five loci, we detected no significant phenotypic variation among lethal alleles. We infer that each of the mutant alleles at these two loci cause expression of the null activity phenotype. However, for the three other loci, we did detect significant phenotypic variation among lethal alleles. In fact, one of the mutant alleles at each of these three loci causes no detectable imaginal disc defect. This demonstrates that attempting to assess the developmental role of a gene by studying a single mutant allele may lead to erroneous conclusions. As a byproduct of the mutagenesis procedure, we have isolated two dominant, cold-sensitive mutants.     

Complementation Analysis of Methyl Methane-Sulfonate-Induced Recessive Lethal Mutations in the Zeste-White Region of the X Chromosome of DROSOPHILA MELANOGASTER

Recessive lethan mutations in the 3A1 to 3C2 region of the X-chromosome of Drosophila melanogaster were detected in 113 of 33,544 sperm treated by feeding 5 mM methyl methanesulfonate in 1% sucrose for 22 hours. Seven of the 113 lethans were sterile, leaving 106 for analysis by complementation tests. With only one exception, these mutants were found to have lesions restricted to single loci. One of these single-site mutations was in qt, 2 in tko, 18 in zw-1, 12 in zw-8, 6 in zw-4, 3 in zw-10, 3 in zw-13, 21 in zw-2, 7 in zw-3, 5 in zw-6, 6 in zw-12, 1 in zw-7, 12 in zw-5, 5 in zw-11, and 3 in zw-9. One of the lethals, m69, was non-complementary to two adjacent loci, zw-2 and zw-3, possible indicating a deletion encompassing two loci. The results confirm that there are at least 15 recessive lethal loci in the region and are consistent with the hypothesis of LIM and SYNDER (1968 and 1974) that inability of monofunctional alkylating chemicals to induce deletion associated mutations is a characteristic of the compounds.     

Gene Flow in Natural Populations of DROSOPHILA MELANOGASTER with Special Reference to Lethal Allelism Rates and Protein Variation

A simultaneous survey of 14 protein loci, together with frequencies and within- and between-population allelism rates of lethal chromosomes, was carried out in five (four Japanese and one Korean) natural populations and one cage population of Drosophila melanogaster. It was found that lethal allelism rates decrease rapidly as geographic distance between two populations increases, while variation at protein loci shows a remarkable similarity over all populations examined. These findings suggest that there are very high levels of gene flow in these natural populations and that selection at protein loci which can maintain substantial geographic variation, if present, is overshadowed by gene flow. There is no indication that invasion of D. melanogaster to the Far East occurred so recently that the frequencies of lethal chromosomes are still in nonequilibrium.     

Recovery and Characterization of Temperature-Sensitive Mutations Affecting Adult Viability in DROSOPHILA MELANOGASTER

Temperature-sensitive (ts) autonomous cell-lethal mutations have been used extensively to study important developmental phenomena, such as pattern formation, in Drosophila. Their utility would be enhanced considerably if it were possible to establish which cell type is primarily affected by each lesion. To facilitate such an approach we have isolated and characterized 21 EMS- induced X-linked adult-lethal (adl) mutants, 16 of which are ts. Most of these lesions also elicit ts lethal effects during preimaginal development. They represent 19 different loci distributed randomly along the X chromosome. The general properties of these mutations are described. In addition, results of an in-depth analysis (focus mapping and, in some cases, temperature shift and heat-pulse studies) of four strains, adl-1^ts1, sesE, adl-2^{ts 1} and rex are reported. Two major temperature-sensitive periods (TSPs) of adl-1 lethality were resolved: one during the second half of embryogenesis and the other coinciding with pupariation. Mosaic analysis revealed separate mesodermal foci for leg paralysis. Developmental analysis of adl-1 embryos suggest that the adl-1 product may be required for maintenance of muscle tissue. Two discrete TSPs of sesE lethality exist: one during the second instar and the other extending from late third instar to early pupation. Mosaic analysis of sesE lethality resolved a pair of neural foci, each of which appears to incorporate three separate foci for leg paralysis. Mosaic analysis of adl-2^{ts 1} revealed the existence of paired lethal foci that appear to map to the vicinity of the subesophageal ganglion. Analysis of rex mosaics resolved separate mesodermal foci for leg paralysis.