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Y N Jan

Publications and source records attributed to Y N Jan.

At least 199 records · Page 11Linked to original sources

Interactions between heterologous helix-loop-helix proteins generate complexes that bind specifically to a common DNA sequence.

A DNA binding and dimerization motif, with apparent amphipathic helices (the HLH motif), has recently been identified in various proteins, including two that bind to immunoglobulin enhancers (E12 and E47). We show here that various HLH proteins can bind as apparent heterodimers to a single DNA motif and also, albeit usually more weakly, as apparent homodimers. The HLH domain can mediate heterodimer formation between either daughterless, E12, or E47 (Class A) and achaete-scute T3 or MyoD (Class B) to form proteins with high affinity for the kappa E2 site in the immunoglobulin kappa chain enhancer. The achaete-scute T3 and MyoD proteins do not form kappa E2-binding heterodimers together, and no active complex with N-myc was evident. The formation of a heterodimer between the daughterless and achaete-scute T3 products may explain the similar phenotypes of mutants at these two loci and the genetic interactions between them. A role of E12 and E47 in mammalian development, analogous to that of daughterless in Drosophila, is likely.

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numb, a gene required in determination of cell fate during sensory organ formation in Drosophila embryos.

Neurons and support cells of each sensory organ in Drosophila embryos are most likely derived from a single precursor cell. This cell lineage is affected in numb mutants. Morphological alterations of sensory structures, as well as changes in the number of cells expressing cell type-specific markers, indicate that sensory neurons in numb mutant embryos are transformed into lineage-related nonneuronal support cells. Thus the numb gene controls the fate of progeny derived from sensory organ precursors. The numb gene has been isolated by the plasmid rescue method. The structure of its predicted product is discussed.

Amino Acid Sequence↗

Neurogenesis of the peripheral nervous system in Drosophila embryos: DNA replication patterns and cell lineages.

Cell lineages that give rise to the PNS were studied using the thymidine analog 5-bromo-2'-deoxyuridine (BrdU) to visualized DNA replication immunocytochemically. The precursors of the PNS in the body segments of Drosophila embryos replicate their DNA in a spatially and temporally stereotyped pattern. The sequence of DNA replication within developing sensory organs suggests particular lineage relationships of the cells that constitute a sensory organ, i.e., neuron and associated support cells. In embryos that are mutant for the achaete-scute complex or daughterless, in which most or all of the PNS is missing, no BrdU-labeled cells were found in the appropriate regions, suggesting that these PNS precursors either do not form or fail to replicate. Thus, the BrdU technique allows determination as to whether a mutation affects the PNS precursors or terminal differentiation.

Animals↗

daughterless, a Drosophila gene essential for both neurogenesis and sex determination, has sequence similarities to myc and the achaete-scute complex.

daughterless (da) has multiple functions in Drosophila embryonic development: maternal da activity is necessary for proper sex determination, and zygotic da activity is necessary for formation of the peripheral nervous system. We have cloned the region containing da and have found that five recessive lethal da mutations map to a single transcription unit. A predicted protein product of this transcription unit has sequence similarities with the oncogene myc, with the gene MyoD1, which is involved in myoblast determination, and with the Drosophila achaete-scute complex, which is involved in neuronal precursor determination. The role of da as a gene controlling cell determination in multiple developmental pathways is discussed.

Amino Acid Sequence↗

A protein component of Drosophila polar granules is encoded by vasa and has extensive sequence similarity to ATP-dependent helicases.

Determinants of pole cells, which are precursors of the germ line, are provided maternally and are localized to the posterior pole of the Drosophila egg, as are polar granules. It has been hypothesized that certain RNA molecules associated with polar granules may be necessary for pole cell determination. Using a monoclonal antibody (Mab46F11) against polar granules, we have cloned the gene for one of their components. This gene turns out to be vasa, which is required maternally for the formation of polar granules and germ cells. This polar granule component shows significant sequence similarity to eIF-4A, a translation initiation factor that binds to mRNA, and to other helicases.

Adenosine Triphosphate↗

Identification and characterization of a neuron-specific nuclear antigen in Drosophila.

An antigen found only in neuronal nuclei of Drosophila melanogaster is revealed by staining with a monoclonal antibody (Mab44C11). This antigen appears early in development, before neurons show any other signs of antigenic or morphologic differentiation, and persists throughout development. This nuclear staining permits reliable detection of neurons in developmental studies of wild-type and mutant flies. Protein immunoblot analyses and immune precipitation experiments show that the neuronal nuclear antigen is a 50-kilodalton polypeptide.

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Cloning of a probable potassium channel gene from mouse brain.

Potassium channels comprise a diverse class of ion channels important for neuronal excitability and plasticity. The recent cloning of the Shaker locus from Drosophila melanogaster has provided a starting point for molecular studies of potassium channels. Predicted Shaker proteins appear to be integral membrane proteins and have a sequence similar to the sequence of the S4 segment of the vertebrate sodium channel, where the S4 segment has been proposed to be the voltage sensor. Expression studies in frog oocytes confirm that Shaker encodes a component of a potassium channel (the A channel) that conducts a fast transient potassium current. Here we report the isolation of complementary DNA clones from the mouse brain, the nucleotide sequences of which predict a protein remarkably similar to the Shaker protein. The strong conservation of the predicted protein sequence in flies and mammals suggests that these mouse clones encode a potassium channel component and that the conserved amino acids may be essential to some aspect of potassium channel function.

Amino Acid Sequence↗

Expression of functional potassium channels from Shaker cDNA in Xenopus oocytes.

The Shaker gene of Drosophila melanogaster encodes a potassium-selective ion channel, the 'A' channel, or one of its subunits. A single Shaker messenger RNA species suffices to direct the synthesis of functional A channels in Xenopus oocytes. Physiological characteristics of the A currents induced by two different mRNA species are compared.

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A carbohydrate epitope expressed uniquely on the cell surface of Drosophila neurons is altered in the mutant nac (neurally altered carbohydrate).

Antibodies against horseradish peroxidase (anti-HRP) recognize neural specific cell surface antigens in Drosophila and other insects. The nature of these antigens was investigated in Drosophila and found to include a complex set of developmentally regulated proteins. Their common epitope appears to be a carbohydrate that shares features with the sugar moiety of pineapple stem bromelain, a plant glycoprotein whose carbohydrate structure has been determined. A mutation was identified that eliminates staining by the antibody in imaginal and adult neural tissue. Tissue specific glycoconjugates, although widespread in the animal kingdom, are little understood. This mutation provides a unique opportunity to address the consequences of altering a neural specific carbohydrate moiety in an otherwise intact and behaving animal. The mutation maps to 84F. A second mutation, contained on the third chromosome balancer, TM3, eliminates anti-HRP staining in embryos. These mutations appear to be separate genes.

Animals↗

Four cDNA clones from the Shaker locus of Drosophila induce kinetically distinct A-type potassium currents in Xenopus oocytes.

The Shaker gene encodes the A-channel of larval and pupal muscle, or one or more of its subunits. Alternative splicing produces messages for several different proteins; two mRNA species have previously been shown to induce the expression of A-currents in Xenopus oocytes. Two additional mRNAs have now been tested and found to produce A-currents in oocytes. The four currents differ in kinetics of inactivation, indicating that the Shaker products may contribute to kinetic diversity in A-channels of the fly and that sequences in both the amino- and carboxy-terminal regions are important for inactivation.

Animals↗

The maternal sex determination gene daughterless has zygotic activity necessary for the formation of peripheral neurons in Drosophila.

The daughterless (da) gene is known to have separate maternal and zygotic functions: Maternally supplied daughterless activity is required for proper sex determination and dosage compensation in female embryos, whereas loss of zygotically supplied da+ activity causes embryonic lethality in both male and female embryos. We have found that the zygotic da+ activity is necessary for neural development: The use of neuron-specific antibodies and beta-galactosidase-marked X chromosomes has revealed that in both male and female embryos deletions or strong mutations of the da gene remove all peripheral neurons and associated sensory structures without disrupting the epithelium from which they derive. Partial da+ function causes partial removal of peripheral neurons. Our results indicate that da+ is required for the formation of peripheral neurons and their associated sensory structures.

Animals↗

Identification of a component of Drosophila polar granules.

Information necessary for the formation of pole cells, precursors of the germ line, is provided maternally and localized to the posterior pole of the Drosophila egg. The maternal origin and posterior localization of polar granules suggest that they may be associated with pole cell determinants. We have generated an antibody (Mab46F11) against polar granules. In oocytes and early embryos, the Mab46F11 antigen is sharply localized to the posterior embryonic pole. In pole cells, it becomes associated with nuclear bodies within, and nuage around, the nucleus. Immunoreactivity remains associated with cells of the germ line throughout the life cycle of both males and females. This antibody recognizes a 72-74 X 10(3) Mr protein and is useful both as a pole lineage marker and in biochemical studies of polar granules.

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Transformation of sensory organs by mutations of the cut locus of D. melanogaster.

The identities of two types of sensory organs in the body wall of Drosophila, namely the external sensory organs and the chordotonal organs, are under genetic control. Embryonic lethal mutations in the cut gene complex transform the external sensory organs into chordotonal organs. The neurons, as well as the support cells forming the external sensory structures, change their morphological and antigenic characteristics to those of chordotonal organs, providing genetic evidence that these two types of sensory organs are homologous. Similar transformations of external sensory organs are observed in adult mosaic flies. Analysis of mosaic larvae and flies suggests that the cut gene function is required either in or near external sensory organs in order for them to acquire their correct identity.

Animals↗

Cloning of genomic and complementary DNA from Shaker, a putative potassium channel gene from Drosophila.

On the basis of electrophysiological analysis of Shaker mutants, the Shaker locus of Drosophila melanogaster has been proposed to encode a structural component of a voltage-dependent potassium channel, the A channel. Unlike sodium channels, acetylcholine receptors, and calcium channels, K+ channels have not been purified biochemically. To facilitate biochemical studies of a K+ channel, genomic DNA from the Shaker locus has been cloned. Rearrangements in five Shaker mutants have been mapped to a 60-kilobase segment of the genome. Four complementary DNA clones have been analyzed. These clones indicate that the Shaker gene contains multiple exons distributed over at least 65 kilobases of genomic DNA in the region where the mutations mapped. Furthermore, the gene may produce several classes of alternatively spliced transcripts. Two of the complementary DNA clones have been sequenced and their sequences support the hypothesis that Shaker encodes a component of a K+ channel.

Animals↗

Sequence of a probable potassium channel component encoded at Shaker locus of Drosophila.

Potassium currents are crucial for the repolarization of electrically excitable membranes, a role that makes potassium channels a target for physiological modifications that alter synaptic efficacy. The Shaker locus of Drosophila is thought to encode a K+ channel. The sequence of two complementary DNA clones from the Shaker locus is reported here. The sequence predicts an integral membrane protein of 70,200 daltons containing seven potential membrane-spanning sequences. In addition, the predicted protein is homologous to the vertebrate sodium channel in a region previously proposed to be involved in the voltage-dependent activation of the Na+ channel. These results support the hypothesis that Shaker encodes a structural component of a voltage-dependent K+ channel and suggest a conserved mechanism for voltage activation.

Animals↗