Functional gene cassettes in development.
Explore the source record for details and available documents.
Animals↗
Biomedical subjects
L Y Jan
Publications and source records attributed to L Y Jan.
Presynaptic A-current based on heteromultimeric K+ channels detected in vivo.
A wide variety of voltage-gated K+ channels are involved in the regulation of neuronal excitability and synaptic transmission. Their heterogeneity arises in part from the large number of genes encoding different K+ channel subunits (reviewed in ref. 1). In addition, heterologous expression studies indicate that assembly of distinct subunits into heteromultimeric channels may contribute further to K+ channel diversity. A question has been whether heteromeric K+ channels actually form in vivo, and if so, whether specific combinations of subunits could account for major K+ currents identified in neurons. We present here biochemical evidence that Kv1.4 and Kv1.2, two K+ channel subunits of the Shaker subfamily, co-assemble in rat brain. The Kv1.4/Kv1.2 heteromultimer combines features of both parent subunits, resulting in an A-type K+ channel. Immunocytochemical evidence suggests that the heteromultimers are localized in axons and nerve terminals. We propose that Kv1.4/Kv1.2 heteromultimers may form the molecular basis of a presynaptic A-type K+ channel involved in the regulation of neurotransmitter release.
Amino Acid Sequence↗
Primary structure and functional expression of a rat G-protein-coupled muscarinic potassium channel.
Parasympathetic nerve stimulation causes slowing of the heart rate by activation of muscarinic receptors and the subsequent opening of muscarinic K+ channels in the sinoatrial node and atrium. This inwardly rectifying K+ channel is coupled directly with G protein. Based on sequence homology with cloned inwardly rectifying K+ channels, ROMK1 (ref. 11) and IRK1 (ref. 12), we have isolated a complementary DNA for a G-protein-coupled inwardly rectifying K+ channel (GIRK1) from rat heart. The GIRK1 channel probably corresponds to the muscarinic K+ channel because (1) its functional properties resemble those of the atrial muscarinic K+ channel and (2) its messenger RNA is much more abundant in the atrium than in the ventricle. In addition, GIRK1 mRNA is expressed not only in the heart but also in the brain.
Amino Acid Sequence↗
atonal is a proneural gene that directs chordotonal organ formation in the Drosophila peripheral nervous system.
In the Drosophila peripheral nervous system, proneural genes of the achaete-scute complex (ASC) are required for formation of the precursors of external sense organs but not of chordotonal organs. We report the isolation of a gene, atonal (ato), with evidence that it is a proneural gene for the formation of chordotonal organs. This gene is expressed in the proneural clusters and sense organ precursors that give rise to the embryonic and adult chordotonal, but not external sense, organs. Chordotonal organs are eliminated in embryos carrying chromosomal deficiencies that remove ato. Like the ASC products, ato protein contains a basic-helix-loop-helix region and heterodimerizes with daughterless protein to bind to E boxes. Moreover, ectopic expression of ato promotes the formation of extra sense organs. Despite similar proneural properties, we find that ectopic expression of the ASC genes promotes external sense organ formation exclusively, whereas ato promotes chordotonal organ formation preferentially. Thus, proneural genes are major determinants of neuronal identity.
Amino Acid Sequence↗
Spatially localized rhomboid is required for establishment of the dorsal-ventral axis in Drosophila oogenesis.
The establishment of dorsal-ventral asymmetry of the Drosophila embryo requires a group of genes that act maternally. None of the previously identified dorsal-ventral axis genes are known to produce asymmetrically localized gene products during oogenesis. We show that rhomboid (rho), a novel member of this group, encodes a protein that is localized on the apical surface of the dorsal-anterior follicle cells surrounding the oocyte. Loss of rho function causes ventralization of the eggshell and the embryo, whereas ectopic expression leads to dorsalization of both structures. Thus, spatially restricted rho is necessary and sufficient for dorsal-ventral axis formation. We propose, based on these observations and double mutant experiments, that the spatially restricted rho protein leads to selective activation of the epidermal growth factor receptor in the dorsal follicle cells and subsequently the specification of the dorsal follicle cells.
Animals↗
Primary structure and functional expression of a mouse inward rectifier potassium channel.
A complementary DNA encoding an inward rectifier K+ channel (IRK1) was isolated from a mouse macrophage cell line by expression cloning. This channel conducts inward K+ current below the K+ equilibrium potential but passes little outward K+ current. The IRK1 channel contains only two putative transmembrane segments per subunit and corresponds to the inner core structure of voltage-gated K+ channels. The IRK1 channel and an ATP-regulated K+ channel show extensive sequence similarity and constitute a new superfamily.
Amino Acid Sequence↗
A new factor related to TATA-binding protein has highly restricted expression patterns in Drosophila.
The TATA-binding protein TBP is necessary for the transcription of eukaryotic genes. Multi-protein complexes formed by TBP and different TBP-associated factors are involved in the initiation of transcription by polymerases I and II, and probably III as well. During the formation of an active initiation complex, TBP makes specific contacts with other proteins, for example TFIIB and RNA polymerase II (refs 2-4). Here we describe the cloning and characterization of a Drosophila gene product with considerable sequence similarity to TBP and a highly restricted expression pattern in the embryo. This TBP-related factor is a DNA-binding protein but is not likely to be a basal transcription factor. Our results suggest that TBP-related factor is a sequence-specific transcription factor that shares the DNA-binding properties of TBP.
Animals↗
The Drosophila neurogenic gene neuralized encodes a novel protein and is expressed in precursors of larval and adult neurons.
Explore the source record for details and available documents.
Amino Acid Sequence↗
Molecular basis of K+ channel inactivation gating.
The gating of many K+, Na+ and Ca++ channels is driven by changes in membrane potential. Part of the gating mechanism, the voltage sensing S4, a proposed transmembrane segment, has been identified. Movement in the membrane electric field of the charged S4 is thought to precede the opening and closing of the activation gate. The physical basis of the conformational changes involved in gating has yet to be elucidated. Here, we discuss a domain that appears to lie at the cytoplasmic mouth of K+ channels and to form a receptor for the inactivation gate. We examine the possibility that a) the physical attachment of this receptor/mouth to the S4 allows inactivation to be coupled to the voltage dependent conformational changes that open the channel and b) explains the immobilization of gating charge by inactivation. We also address the physiological ramifications of such structural coupling.
Amino Acid Sequence↗
The S4-S5 loop contributes to the ion-selective pore of potassium channels.
Mutagenesis experiments on voltage-gated K+ channels have suggested that the ion-selective pore is comprised mostly of H5 segments. To see whether regions outside of the H5 segment might also contribute to the pore structure, we have studied the effect of single amino acid substitutions in the segment that connects the S4 and S5 putative transmembrane segments (S4-S5 loop) on various permeation properties of Shaker K+ channels. Mutations in the S4-S5 loop alter the Rb+ selectivity, the single-channel K+ and Rb+ conductances, and the sensitivity to open channel block produced by intracellular tetraethylammonium ion, Ba2+, and Mg2+. The block of Shaker K+ channels by intracellular Mg2+ is surprising, but is reminiscent of the internal Mg2+ blockade of inward rectifier K+ channels. The results suggest that the S4-S5 loop constitutes part of the ion-selective pore. Thus, the S4-S5 loop and the H5 segment are likely to contribute to the long pore characteristic of voltage-gated K+ channels.
Amino Acid Sequence↗
Cell interactions and gene interactions in peripheral neurogenesis.
Explore the source record for details and available documents.
Animals↗
A rat gene with sequence homology to the Drosophila gene hairy is rapidly induced by growth factors known to influence neuronal differentiation.
Several genes encoding transcription factors with a helix-loop-helix (HLH) motif are involved in the early process of neural development in Drosophila spp. We report the isolation from the rat a homolog of one of these genes, called hairy. The rat-hairy-like (RHL) gene is expressed early during embryogenesis. In contrast to the restricted expression of hairy mRNA in Drosophila spp., however, the mRNA encoded by RHL is detectable in all tissues examined. Stimulation of PC12 pheochromocytoma cells by nerve growth factor, basis fibroblast growth factor, or epidermal growth factor or of Rat-1 fibroblasts by epidermal growth factor causes a rapid and transient induction of the RHL gene. Thus, RHL acts as an immediate-early gene that can potentially transduce growth factor signals during the development of the mammalian embryo.
Amino Acid Sequence↗
Specifying the path of the intersegmental nerve of the Drosophila embryo: a role for Delta and Notch.
The intersegmental nerve (ISN) of the Drosophila embryo follows a reproducible course near the anterior border of each segment. Based on the experiments reported here, we suggest that growth of the axons constituting the nerve is guided, in part, by the transmembrane proteins Delta and Notch. In particular, we suggest that expression of Delta protein on a branch of the trachea provides a path for the nerve through the lateral part of the embryo, and that the growing axons use the Notch protein on their surfaces to recognize this path. Consistent with this idea, we show that disruption of the trachea abolishes the ability of the ISN to extend through this part of the embryonic periphery. Finally, we argue that the same regulatory network that directs these peripheral axons also specifies the trajectory of part of the axonal scaffold of the central nervous system.
Animals↗
Postembryonic patterns of expression of cut, a locus regulating sensory organ identity in Drosophila.
The cut locus is both necessary and sufficient to specify the identity of a class of sensory organs in Drosophila embryos. It is also expressed in and required for the development of a number of other embryonic tissues, such as the central nervous system, the Malpighian tubules and the tracheal system. We here describe the expression of cut in the precursors of adult sensory organs. We also show that cut is expressed in cells of the prospective wing margin and correlate the wing margin phenotype caused by two cut mutations with altered cut expression patterns. Finally, we observe cut-expressing cells in other adult tissues, including Malpighian tubules, muscles, the central nervous system and ovarian follicle cells.
Animals↗
pipsqueak, an early acting member of the posterior group of genes, affects vasa level and germ cell-somatic cell interaction in the developing egg chamber.
We have identified a new member of the posterior group of genes, which we call pipsqueak. We show that pipsqueak acts after the establishment of the oskar posterior anchor but before the localization of vasa protein during oogenesis. Characterization of multiple alleles at the pipsqueak locus shows that pipsqueak, like vasa, is required for early stages of oogenesis, including but not limited to formation of the egg chamber and progression through Stage 6 of oogenesis. Genetic interaction studies suggest that pipsqueak acts at least partially through vasa; molecular studies indicate that pipsqueak affects vasa level in the ovary. We compare vasa and pipsqueak mutant phenotypes in order to determine whether pipsqueak acts solely through vasa, and present a model for the role of pipsqueak in posterior pattern formation.
Animals↗
asense is a Drosophila neural precursor gene and is capable of initiating sense organ formation.
Neural precursor cells in Drosophila arise from the ectoderm in the embryo and from imaginal disc epithelia in the larva. In both cases, this process requires daughterless and the proneural genes achaete, scute and lethal-of-scute of the achaete-scute complex. These genes encode basic helix-loop-helix proteins, which are nuclear transcription factors, as does the asense gene of the achaete-scute complex. Our studies suggest that asense is a neural precursor gene, rather than a proneural gene. Unlike the proneural achaete-scute gene products, the asense RNA and protein are found in the neural precursor during its formation, but not in the proneural cluster of cells that gives rise to the neural precursor cell. Also, asense expression persists longer during neural precursor development than the proneural gene products; it is still expressed after the first division of the neural precursor. Moreover, asense is likely to be downstream of the proneural genes, because (1) asense expression is affected in proneural and neurogenic mutant backgrounds, (2) ectopic expression of asense protein with an intact DNA-binding domain bypasses the requirement for achaete and scute in the formation of imaginal sense organs. We further note that asense ectopic expression is capable of initiating the sense organ fate in cells that do not normally require the action of asense. Our studies therefore serve as a cautionary note for the inference of normal gene function based on the gain-of-function phenotype after ectopic expression.
Amino Acid Sequence↗
The regulation and function of the helix-loop-helix gene, asense, in Drosophila neural precursors.
asense is a member of the achaete-scute complex (AS-C) of helix-loop-helix genes involved in Drosophila neurogenesis. Unlike the other AS-C members, which are expressed in subsets of the ectodermal areas (proneural clusters) that give rise to neural precursors, asense is one of a number of genes that are specifically expressed in the neural precursors themselves (neural precursor genes). We have identified a mutant asense phenotype that may reflect this later expression pattern. As a step in understanding the determination of neural precursors from the proneural clusters, we have investigated the potential role of the AS-C products as direct transcriptional activators of neural precursor genes by analysing the regulation of asense. Using genomic rescues and asense-lacZ fusion genes, the neural precursor regulatory element has been identified. We show that this element contains binding sites for AS-C/daughterless heterodimers. Delection of these sites reduces the expression from the fusion gene, but significant expression is still achieved, pointing to the existence of other regulators of asense in addition to the AS-C. asense differs from the other AS-C members in its expression pattern, regulation, mutant phenotype and some DNA-binding properities.
Animals↗
deadpan, an essential pan-neural gene encoding an HLH protein, acts as a denominator in Drosophila sex determination.
In Drosophila, sex is determined by the X:A ratio. One major numerator element on the X chromosome is sisterless-b (sis-b), also called scute, which encodes an HLH-type transcription factor. We report here that an essential pan-neural gene, the autosomal HLH gene deadpan (dpn), acts as a denominator element. As revealed by dosage-dependent dominant interactions, males die with too high a ratio of sc+ to dpn+, caused by misexpression of Sex lethal (Sxl) in embryos, and females die with too low a ratio of sc+ to dpn+, because of altered embryonic Sxl expression. In addition, we found that the HLH gene extramacrochaetae (emc), like daughterless (da), is needed maternally for proper communication of the X:A ratio, thus supporting the idea that a set of HLH genes comprises a functional cassette that makes a sensitive and stable genetic switch used in both neural determination and sex determination.
Animals↗