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K D Konrad

Publications and source records attributed to K D Konrad.

11 recordsLinked to original sources
PubMed1998

The gastrulation defective gene of Drosophila melanogaster is a member of the serine protease superfamily.

The establishment of dorsal-ventral polarity in the oocyte involves two sets of genes. One set belongs to the gurken-torpedo signaling pathway and affects the development of the egg chorion as well as the polarity of the embryo. The second set of genes affects only the dorsal-ventral polarity of the embryo but not the eggshell. gastrulation defective is one of the earliest acting of this second set of maternally required genes. We have cloned and characterized the gastrulation defective gene and determined that it encodes a protein structurally related to the serine protease superfamily, which also includes the Snake, Easter, and Nudel proteins. These data provide additional support for the involvement of a protease cascade in generating an asymmetric signal (i.e., asymmetric Spätzle activity) during establishment of dorsal-ventral polarity in the Drosophila embryo.

Amino Acid Sequence↗
PubMed1994

Dorsal midline fate in Drosophila embryos requires twisted gastrulation, a gene encoding a secreted protein related to human connective tissue growth factor.

The twisted gastrulation (tsg) gene is one of seven known zygotic genes that specify the fate of dorsal cells in Drosophila embryos. Mutations in these genes cause at least some of the cells on the dorsal half of the embryo to adopt more ventral cell fates leading to the proposal that most of these genes participate in establishing, maintaining, or modulating a gradient of a single signaling molecule DECAPENTAPLEGIC (DPP). We have examined the effects of tsg mutations on the development of cuticule elements, expression of a region specific enhancer trap, and patterns of mitotic domains. Mutations of tsg only affect the fate of a narrow strip of dorsal midline cells and do not affect dorsal ectoderm cells. However, the pattern of tsg expression is not coincident with the territories affected by tsg mutations. Structural analysis of the tsg gene reveals features of a secreted protein suggesting an extracellular site of action. The TSG protein bears a weak resemblance to human connective tissue growth factor (CTGF), a TGF-beta-induced protein. We propose that dorsal midline cell fate is specified by the combination of both a TSG and a DPP signal to which the dorsal midline cells are uniquely competent to respond.

Algorithms↗
PubMed1993

Vitelline membrane biogenesis in Drosophila requires the activity of the alpha-methyl dopa hypersensitive gene (I(2)amd) in both the germline and follicle cells.

The vitelline membrane of Drosophila eggs is composed of a family of proteins which are cross linked into an insoluble matrix with an overlying waxy layer that prevents desiccation. We present here three sets of experiments which show that integrity of the vitelline membrane requires the activity of the alpha methyl dopa hypersensitive (I(2)amdH or amd) gene in both egg chambers and follicle cells. We show that loss of amd activity either by dietary administration of inhibitors or in genetic mosaics of either the germ line or follicle cells leads to production of defective vitelline membranes and by in situ hybridization, that amd is expressed in both nurse cells and follicle cells. The amd gene product is the first non-structural protein gene described whose activity is required for vitelline membrane biosynthesis. Given its unique role in insects and its demonstrated sensitivity to dietary inhibitors, the amd gene product poses an attractive target for insect control.

Animals↗
PubMed1992

Drosophila homolog of the human S6 ribosomal protein is required for tumor suppression in the hematopoietic system.

The tumor suppressor gene lethal(1)aberrant immune response 8 (air8) of Drosophila melanogaster encodes a homolog of the human S6 ribosomal protein. P element insertions that prevent expression of this gene cause overgrowth of the lymph glands (the hematopoietic organs), abnormal blood cell differentiation, and melanotic tumor formation. They also cause delayed development, inhibit growth of most of the larval organs, and lead to larval lethality. Mitotic recombination experiments indicate that the normal S6 gene is required for clone survival in the germ line and imaginal discs. The S6 gene produces a 1.1-kilobase transcript that is abundant throughout development in wild-type animals and in revertants derived from the insertional mutants but is barely detectable in the mutant larvae. cDNAs corresponding to this transcript show a 248-amino acid open reading frame with 75.4% identity and 94.8% similarity to both human and rat S6 ribosomal protein sequences. The results reveal a regulatory function of this ribosomal protein in the hematopoietic system of Drosophila that may be related to its developmentally regulated phosphorylation.

Amino Acid Sequence↗
PubMed1992

A single-fluor approach to DNA sequence determination using high performance capillary electrophoresis.

The Tabor and Richardson strategy for enzymatic chain termination sequencing of DNA using relative peak intensity has been adapted to high performance capillary gel electrophoresis with laser induced fluorescence detection. This approach to DNA sequencing involves the use of only a single fluor and results in significant reduction in the time required to determine a DNA sequence without the use of highly complicated and expensive instrumentation. We present a modification of the Tabor and Richardson approach employing two reactions, each containing complementary mixtures of only three ddNTP's in the concentration ratio 4:2:1. The DNA sequence is determined by relative peak height and by assigning the missing ddNTP to "gaps" between the peaks. The use of only three terminators/reaction simplifies the software task of differentiating between the termination types and makes more efficient use of the available dynamic range. Both complementary mixes generate complete sequence information and the two data files are combined in order to make a more confident sequence call. This process helps to eliminate errors caused by occasional non-uniform incorporation of ddNTP's or false terminations and also alleviates some of the difficulty associated with reading through compressed regions of the electropherogram.

Base Sequence↗
PubMed1988

Developmental genetics of the gastrulation defective locus in Drosophila melanogaster.

The fs(1)gastrulation defective (dg) locus is one of the dorsal-group genes of Drosophila. Maternal expression of this gene is required for gastrulation movements and the differentiation of structures along the embryonic dorso-ventral axis. Twelve alleles of gd displayed a complex pattern of complementation, suggesting a direct interaction between subunits of a multimeric protein. Essential expression of the gd locus was strictly maternal with no zygotic contribution by the paternally derived allele. Clonal analysis revealed that expression of the gd locus was required in the germ line and that extreme dorsalization represented the null gd phenotype. Temperature-sensitive (ts) alleles displayed a ts period that included the last 4-5 hr of oogenesis and the first 1.5-2 hr of embryogenesis. Eggs from one ts allelic combination displayed reduced hatching when retained in the ovary at permissive temperatures, suggesting the loss of a labile egg component. This lability may also be responsible for the variable phenotypes displayed by offspring from individual females.

Alleles↗
PubMed1987

Developmental expression and spatial distribution of dopa decarboxylase in Drosophila.

Regulation of the dopa decarboxylase gene of Drosophila has been studied at the genetic and molecular levels. Here we report a direct assay for the tissue and temporal regulation of Ddc. A dopa decarboxylase (DDC) peptide was obtained by bacterial expression of a portion of the DDC gene in a pUC plasmid. Antisera raised against this biologically purified DDC peptide react specifically with Drosophila DDC in histological preparations and protein blots. The levels of DDC cross-reacting material closely parallel the levels of enzyme activity observed during development, indicating that DDC is degraded during periods of declining activity. We find that DDC is expressed in only two tissues, namely, the epidermis and the nervous system of the larva and adult. Epidermal DDC was found within the epidermal cells and was not detected in the overlying cuticle. DDC-containing neurons were observed in the central as well as in the visceral nervous system. Paired and unpaired midline neurons in the ventral ganglia are arranged in a segmental pattern. A subset of the DDC-positive neurons appears to correlate with the serotonin-positive neurons suggesting that the others are producing only dopamine. We find that the DDC activity associated with the proventriculus and ovary is due to the presence of DDC in the stomatogastric and caudal system neurons specifically associated with those structures.

Animals↗
PubMedDate not supplied

Contribution of secondary structure to DNA mobility in capillary gels.

Intramolecular base pairing at the 3' end of single-stranded DNA molecules can cause increased electrophoretic mobility. In DNA sequencing separations this can result in compressions, where multiple oligomers differing in length by one base comigrate, which complicate sequence analysis. Using a novel approach to the application of capillary electrophoresis to DNA sequencing, incorporation of formamide into capillary gels, different separation field strengths, and external heating of the capillary are examined for their ability to resolve compressions. The identity of the 3' terminal nucleotide has also been observed to influence oligomer mobility in cross-linked acrylamide slab gels. We utilize automated data collection from DNA sequencing separations to show that a similar effect is also present in linear acrylamide capillary gels run at high field strengths. We also demonstrate the ability of capillary electrophoresis to separate synthetic oligos of the same length but differing in composition by one base, or of the same base composition but different sequence.

Base Sequence↗