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Drosophila embryos homozygous for strong mutations in each of the segment-polarity genes wingless (wg), engrailed (en), naked (nkd) and patched (ptc) form a larval cuticle in which there is a deletion in every segment. The mutant embryos normally fail to hatch but by in vivo culture we were able to show which could produce adult structures. Cultured wg− embryos did not produce any adult structures. Cultured en− embryos produced eye-antennal derivatives and rarely produced partial thoracic structures. nkd− and ptc− embryos produced eye-antennal and thoracic derivatives. The nkd− and ptc− thoracic imaginal discs developed with an abnormal morphology and abnormal pattern of en-expression. Our findings are consistent with the idea that the thoracic imaginal discs derive from two adjacent groups of cells that express wg and en respectively in the embryo.

We demonstrate the role of the segment polarity gene patched (ptc) in patterning in the cuticle of the adult fly. Genetic mosaics of a lethal allele of patched show that the contribution of patched varies in a position-specific manner, defining three regions in the wing where ptc clones, respectively, behave as wild-type cells, affect vein formation, or are rarely recovered. Analysis of twin clones demonstrates that the reduced clone frequency results from a proliferation failure or cell loss. In the region where clones upset venation, they autonomously fail to form veins and also non-autonomously induce ectopic veins in adjacent wild-type cells. In heteroallelic combinations with lethal alleles, two viable alleles produce distinct phenotypes: (1) loss of structures and mirror-image duplications in the region where patched clones fail to proliferate; (2) vein abnormalities in the anterior compartment. We propose that these differ-enees reflect independently mutable functions within the gene. We show the pattern of patched transcription in the developing imaginal wing disc in relation to the expression of certain other reporter genes using a novel double-labelling method combining non-radioactive detection of in situ hybridization with β-galactosidase detection. The patched transcript is present throughout the anterior compartment, with a stripe of maximal intensity along the A/P compartment border extending into the posterior compartment. We propose that the patched product is a component of a cell-to-cell positionsignalling mechanism, a proposal consistent with the predicted structure of the patched protein.

We have examined the developmental consequences for larval and imaginal segmental cuticular structure of a chromosomal translocation involving a breakpoint in the abdominal region of the bithorax complex (BX-C). This complex makes an essential contribution to the development of metameric differences in part of the thorax and in all abdominal segments. The breakpoint is proximal to the most distal (iab-7) homeobox, and results in the translocation to the Y chromosome of the Ultrabithorax (Ubx) and abdominal-A (abd-A) domains. The genotype deficient for the distal part of the complex shows normal function for Ubx and abd-A but has a phenotype typical for severe Abd-B mutations. Conversely, the distal fragment retains a segment identity function which must represent a contribution from Abd-B in parasegments 13 and 14; the latter metamere is wild type, indicating that it does not require the contribution of Ubx or abd-A. We also constructed a genotype comprising the proximal fragment of this translocation together with an overlapping distal fragment of the BX-C derived from Df(3R)Ubx109. It therefore contained all sequences of the BX-C though in the abdominal region the abd-A and Abd-B domains were not adjacent to each other in the chromosome. This genotype was phenotypically normal and demonstrates that DNA sequences in the abd-A and Abd-B regions do not require cis-arrangement for their activity.

SUMMARYIn order to investigate the importance of the nuclear genotype during morphogenesis of the cortex inParamecium aurelia, a search was made for conditional and unconditional mutants affecting cell shape. Ten different mutants were readily recovered following mutagenesis withN-methyl-N-nitro-N-nitrosoguanidine on stock 51s of syngen 4. The detailed pattern of surface units remained unchanged in these mutants but each one can be described as resembling a normal cell twisted about its long axis. Nine of the mutants complement each other and show no linkage. The remaining mutant, a ‘chain former’, has conditional expression, and is only recognizable at 35 °C. The penetrance of this mutant is affected by the food organism used. The conditional mutant also interacts with seven of the other mutants in pairwise homozygous combinations so that in these genotypes it can be classified after growth at 25 °C. The mutants are discussed in relation to the search for genie effects on the cortical pattern.

SUMMARYA genetic analysis was made of theFemale-lethal(Fl) locus ofDrosophila melanogaster. This is anX-linked mutation which causes lethality only in females. Other alleles do not complementFland are either lethal or sterile when homozygous in females. Complementation studies onFlalleles demonstrate that there is no simple ranking of these alleles in terms of severity of phenotypic effect. Dosage manipulation ofFlalleles indicates that the sex-specificity is not a consequence of gene dosage effects. Viability studies on males carryingFlalleles show thatFlalleles have no effect on viability regardless of the presence or absence of aYchromosome. TheFllocus is therefore sex-specific. The hypothesis thatFl+ is involved in the establishment of imaginal phenotypic sex cannot be substantiated on the basis of experiments utilizing sex-change mutations.

The patterning of cells in insect segments requires the exchange of information between cells, which in Drosophila depends on the activity of members of the segment-polarity class of genes. Here we report the molecular characterization of one such gene, patched. We find that patched encodes a large protein with several possible membrane-spanning domains and is expressed in a complex pattern during embryogenesis.

The usefulness in structure/function studies of molybdenum-containing hydroxylases in work with rosy mutant strains of Drosophila melanogaster has been investigated. At least 23 such strains are available, each corresponding to a single known amino acid change in the xanthine dehydrogenase sequence. Sequence comparisons permit identification, with some certainty, of regions associated with the iron-sulphur centres and the pterin molybdenum cofactor of the enzyme. Procedures have been developed and rigorously tested for the assay in gel-filtered extracts of the flies, of different catalytic activities of xanthine dehydrogenase by the use of various oxidizing and reducing substrates. These methods have been applied to 11 different rosy mutant strains that map to different regions of the sequence. All the mutations studied cause characteristic activity changes in the enzyme. In general these are consistent with the accepted assignment of the cofactors to the different domains and with the known reactivities of the molybdenum, flavin and iron-sulphur centres. Most results are interpretable in terms of the mutation affecting electron transfer to or from one redox centre only. The activity data provide evidence that FAD and the NAD+/NADH binding sites are retained in mutants mapping to the flavin domain. Therefore, despite some indications from sequence comparisons, it is concluded that the structure of this domain of xanthine dehydrogenase cannot be directly related to that of other flavoproteins for which structural data are available. The data also indicate that the artificial electron acceptor phenazine methosulphate acts at the iron-sulphur centres and suggest that these centres may not be essential for electron transfer between molybdenum and flavin. The work emphasizes the importance of combined genetic and biochemical study of rosy mutant xanthine dehydrogenase variants in probing the structure and function of enzymes of this class.

The Organization of Heredity . By Kenneth R. Lewis & Bernard John. Pp. ix + 241. (Arnold: London, September 1970.) 70s boards; 35s paper.