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J R Whittle

Publications and source records attributed to J R Whittle.

15 recordsLinked to original sources
PubMed1992

Use of rosy mutant strains of Drosophila melanogaster to probe the structure and function of xanthine dehydrogenase.

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.

Amino Acid Sequence↗
PubMed1991

Establishment of imaginal discs and histoblast nests in Drosophila.

In Drosophila the homeotic genes of the bithorax-complex (BX-C) and Antennapedia-complex (ANT-C) specify the identity of segments. Adult segment primordia are established in the embryo as the histoblast nests of the abdomen and the imaginal discs of the head, thorax and terminalia. We have used a molecular probe for the limb primordia and in vivo culture to describe the nature of the adult primordia in mutants in which the pattern of homeotic gene expression was altered. The results suggest that the histoblast or disc 'mode' of development is initiated by the extended germ band stage through activity of the BX-C and ANT-C and is relatively inflexible thereafter [corrected].

Animals↗
PubMed1990

The Drosophila segment polarity gene patched is involved in a position-signalling mechanism in imaginal discs.

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 differences 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 beta-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 position-signalling mechanism, a proposal consistent with the predicted structure of the patched protein.

Alleles↗
PubMed1990

Pattern formation in imaginal discs.

Imaginal discs are sacs of folded epithelium arising during embryogenesis. They proliferate during the larval instars, and at metamorphosis secrete the adult cuticle, therefore being responsible for the characteristic surface patterning of this insect. Each disc has intrinsic growth controls and their cell lineage shows constraints known as compartments. Spatial patterns emerge through interaction between adjacent cells. Molecular genetic analysis of mutants with changed pattern has implicated transcription factors, secreted, membrane-bound and growth factor related proteins in the position-signalling mechanism. Their accessibility to contemporary cell biological techniques makes imaginal discs a model system for investigating patterning in animal tissues.

Animals↗
PubMed1989

A protein with several possible membrane-spanning domains encoded by the Drosophila segment polarity gene patched.

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.

Alleles↗
PubMed1987

Chromosomal continuity in the abdominal region of the bithorax complex of Drosophila is not essential for its contribution to metameric identity.

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.

Animals↗
PubMed1986

The effect of lethal mutations and deletions within the bithorax complex upon the identity of caudal metameres in the Drosophila embryo.

Mutations and deletions of the abdA and AbdB functions in the bithorax complex of Drosophila melanogaster have been examined for their effect upon the hypodermal derivatives of the caudal segments of the embryo, employing light- and scanning electron microscopy. No cuticular structures located posterior to the denticle belt of abdominal segment 8 are affected in abdA- embryos. Embryos of AbdB- genotype no longer have six of the seven pairs of sense organs present in this region, lack posterior spiracles but instead have sclerotized cuticle and sense organs typical of the head region and a rudimentary extra ventral denticle belt. The anal pads, tuft and sense organ 1 do not require BX-C functions for their specification. We discuss the provenance of these cuticular structures and the domain of function of elements within the bithorax complex in terms of parasegmental metameric units.

Animals↗
PubMed1985

Recessive lethal mutations within the bithorax-complex in Drosophila.

Genetic deficiencies of the bithorax-complex (BX-C) in Drosophila, have been used to recover recessive lethal mutations in this chromosome region following mutagenesis. Complementation analysis separates these lethal mutations into five groups within a smaller deficiency, thought to remove the entire BX-C, and into 20 to the left and 4 to the right of the region. Homozygotes for each of only three groups of lethals, Ubx, abdA and AbdB, produce homoeotic segmental transformations in embryos. The functional domains of abdA and AbdB have been defined by changes in the appearance of larval hypodermal structures and of clones in imaginal tissue. The function abdA is required in all the compartments caudal to the anteroposterior border of abdominal segment 1 up to and including the anterior region of abdominal segment 8, whilst AbdB is required in abdominal segments 5 to 9. One allele of AbdB produces a ninth abdominal setal band and structures characteristic of head segments posterior to A8. Rare adult survivors hemizygous for an AbdB allele have eight abdominal segments in both sexes, and lack genitalia in females. Our findings are discussed in the context of the organisation of genetic functions within the BX-C.

Animals↗
PubMed1979

Replacement of posterior by anterior structures in the Drosophila wing caused by the mutation apterous-blot.

The recessive mutation apterous-blot in Drosophila melanogaster causes replacement of posterior wing structures by anterior ones, with variable penetrance and expressivity. Extreme transformations resemble mirror-image duplicate anterior wings as in the mutant engrailed. Anterior structures in the posterior wing only appear on the dorsal surface. Duplications solely of posterior structures are also seen. Clonal analysis shows that extra cell proliferation occurs in the posterior area but is complete by 108 h after egg deposition. Lineage analysis is consistent with a clonal perpetuation of the transformation. Genetic mosaics to test the cell-autonomy of apterous-blot show that it is not autonomously expressed in clones. The results of lineage analysis, the phenotypes of combinations of apterous-blot with other apterous alleles including a deletion for the locus and with various other homoeotic mutations, are together used to distinguish three alternative modes of action of this mutation. It is concluded that apterous-blot is unlikely to be a selector gene mutation but instead may cause the transformation by an event like transdetermination following a local failure in cell function in the wing disc.

Animals↗