scholarly journals Kinesin-2 transports Orco into the olfactory cilium of Drosophila melanogaster at specific developmental stages

PLoS Genetics ◽  
2021 ◽  
Vol 17 (8) ◽  
pp. e1009752
Author(s):  
Swadhin Chandra Jana ◽  
Priya Dutta ◽  
Akanksha Jain ◽  
Anjusha Singh ◽  
Lavanya Adusumilli ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Outer Segment ◽  
Developmental Stages ◽  
Transmembrane Protein ◽  
Developmental Regulation ◽  
Cell Signalling ◽  
Intraflagellar Transport ◽  
Distal Portion ◽  
Electrophysiological Response ◽  
Associated Proteins

The cilium, the sensing centre for the cell, displays an extensive repertoire of receptors for various cell signalling processes. The dynamic nature of ciliary signalling indicates that the ciliary entry of receptors and associated proteins must be regulated and conditional. To understand this process, we studied the ciliary localisation of the odour-receptor coreceptor (Orco), a seven-pass transmembrane protein essential for insect olfaction. Little is known about when and how Orco gets into the cilia. Here, using Drosophila melanogaster, we show that the bulk of Orco selectively enters the cilia on adult olfactory sensory neurons in two discrete, one-hour intervals after eclosion. A conditional loss of heterotrimeric kinesin-2 during this period reduces the electrophysiological response to odours and affects olfactory behaviour. We further show that Orco binds to the C-terminal tail fragments of the heterotrimeric kinesin-2 motor, which is required to transfer Orco from the ciliary base to the outer segment and maintain within an approximately four-micron stretch at the distal portion of the ciliary outer-segment. The Orco transport was not affected by the loss of critical intraflagellar transport components, IFT172/Oseg2 and IFT88/NompB, respectively, during the adult stage. These results highlight a novel developmental regulation of seven-pass transmembrane receptor transport into the cilia and indicate that ciliary signalling is both developmentally and temporally regulated.

scholarly journals Heterotrimeric kinesin-2 motor subunit, KLP68D, localises Drosophila odour receptor coreceptor in the distal domain of the olfactory cilia

2020 ◽  
Author(s):  
Swadhin C. Jana ◽  
Akanksha Jain ◽  
Priya Dutta ◽  
Anjusha Singh ◽  
Lavanya Adusumilli ◽  
...  
Keyword(s):  
Sensory Neurons ◽  
Outer Segment ◽  
Developmental Stages ◽  
Olfactory Sensory Neurons ◽  
List Type ◽  
Tail Domain ◽  
Specific Domain ◽  
Electrophysiological Response ◽  
Olfactory Cilia ◽  
Specific Regulation

AbstractCiliary localisation of the odour receptor coreceptor (Orco) is essential for insect olfaction. Here, we show that in the Drosophila antenna Orco enters the bipartite cilia expressed on the olfactory sensory neurons in two discrete, one-hour intervals after the adult eclosion. Genetic analyses suggest that the heterotrimeric kinesin-2 is essential for Orco transfer from the base into the cilium. Using in vitro pulldown assay, we show that Orco binds to the C-terminal tail domain of KLP68D, the β-subunit of kinesin-2. Reduced Orco enrichment decreases electrophysiological response to odours and loss of olfactory behaviour. Finally, we show that kinesin-2 function is necessary to compact Orco to an approximately four-micron stretch at the distal portion of the ciliary outer-segment bearing singlet microtubule filaments. Altogether, these results highlight an independent, tissue-specific regulation of Orco entry at specific developmental stages and its localisation to a ciliary subdomain by kinesin-2.Graphical AbstractAuthor SummaryJana, Jain, Dutta et al., show that the odour receptor coreceptor only enters the cilia expressed on olfactory sensory neurons at specified developmental stages requiring heterotrimeric kinesin-2. The motor also helps to localise the coreceptor in a compact, environment-exposed domain at the ciliary outer-segment.Highlights:Odorant receptor coreceptor (Orco) selectively enters the olfactory cilia.Orco localises in a specific domain at the distal segment of the olfactory cilium.Orco/ORx binds to the C-terminal tail domain of the kinesin-2β motor subunit.Orco entry across the transition zone and its positioning require Kinesin-2.

scholarly journals The actin cytoskeleton of Dictyostelium: a story told by mutants

2000 ◽  
Vol 113 (5) ◽  
pp. 759-766 ◽  
Author(s):  
A.A. Noegel ◽  
M. Schleicher
Keyword(s):  
Actin Cytoskeleton ◽  
Binding Proteins ◽  
Developmental Stages ◽  
Temporal Dynamics ◽  
Cell Signalling ◽  
Actin Binding ◽  
Actin Binding Proteins ◽  
Associated Proteins ◽  
Bona Fide ◽  
Cellular Behaviour

Actin-binding proteins are effectors of cell signalling and coordinators of cellular behaviour. Research on the Dictyostelium actin cytoskeleton has focused both on the elucidation of the function of bona fide actin-binding proteins as well as on proteins involved in signalling to the cytoskeleton. A major part of this work is concerned with the analysis of Dictyostelium mutants. The results derived from these investigations have added to our understanding of the role of the actin cytoskeleton in growth and development. Furthermore, the studies have identified several cellular and developmental stages that are particularly sensitive to an unbalanced cytoskeleton. In addition, use of GFP fusion proteins is revealing the spatial and temporal dynamics of interactions between actin-associated proteins and the cytoskeleton.

scholarly journals The role of NbTMP1, a surface protein of sporoplasm, in Nosema bombycis infection

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Shiyi Zheng ◽  
Yukang Huang ◽  
Hongyun Huang ◽  
Bin Yu ◽  
Ni Zhou ◽  
...  
Keyword(s):  
Indirect Immunofluorescence ◽  
Selective Breeding ◽  
Developmental Stages ◽  
Transmembrane Protein ◽  
Surface Protein ◽  
Spore Wall ◽  
Significant Inhibition ◽  
Pcr Analysis ◽  
Immunofluorescence Analysis ◽  
Nosema Bombycis

Abstract Background Nosema bombycis is a unicellular eukaryotic pathogen of the silkworm, Bombyx mori, and is an economic and occupational hazard in the silkworm industry. Because of its long incubation period and horizontal and vertical transmission, it is subject to quarantine measures in sericulture production. The microsporidian life-cycle includes a dormant extracellular phase and intracellular proliferation phase, with the proliferation period being the most active period. This latter period lacks spore wall protection and may be the most susceptible stage for control. Methods In order to find suitable target for the selective breeding of N. bombycis-resistant silkworm strains, we screen highly expressed membrane proteins from the transcriptome data of N. bombycis. The subcellular localization of the candidate protein was verified by Indirect immunofluorescence analysis (IFA) and immunoelectron microscopy (IEM), and its role in N. bombycis proliferation was verified by RNAi. Results The N. bombycis protein (NBO_76g0014) was identified as a transmembrane protein and named NbTMP1. It is homologous with hypothetical proteins NGRA_1734 from Nosema granulosis. NbTMP1 has a transmembrane region of 23 amino acids at the N-terminus. Indirect immunofluorescence analysis (IFA) results suggest that NbTMP1 is secreted on the plasma membrane as the spores develop. Western blot and qRT-PCR analysis showed that NbTMP1 was expressed in all developmental stages of N. bombycis in infected cells and in the silkworm midgut. Downregulation of NbTMP1 expression resulted in significant inhibition of N. bombycis proliferation. Conclusions We confirmed that NbTMP1 is a membrane protein of N. bombycis. Reduction of the transcription level of NbTMP1 significantly inhibited N. bombycis proliferation, and this protein may be a target for the selective breeding of N. bombycis-resistant silkworm strains.

scholarly journals Development and Interrogation of a Transcriptomic Resource for the Giant Triton Snail (Charonia tritonis)

2021 ◽  
Author(s):  
AH Klein ◽  
CA Motti ◽  
AK Hillberg ◽  
T Ventura ◽  
P Thomas-Hall ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Cell Signalling ◽  
Natural Populations ◽  
Olfactory Receptors ◽  
Gill Tissue ◽  
Abundant Species ◽  
Population Outbreaks ◽  
Coral Reef Ecosystems ◽  
Specialist Predators ◽  
G Protein Coupled

AbstractGastropod molluscs are among the most abundant species that inhabit coral reef ecosystems. Many are specialist predators, along with the giant triton snail Charonia tritonis (Linnaeus, 1758) whose diet consists of Acanthaster planci (crown-of-thorns starfish), a corallivore known to consume enormous quantities of reef-building coral. C. tritonis are considered vulnerable due to overexploitation, and a decline in their populations is believed to have contributed to recurring A. planci population outbreaks. Aquaculture is considered one approach that could help restore natural populations of C. tritonis and mitigate coral loss; however, numerous questions remain unanswered regarding their life cycle, including the molecular factors that regulate their reproduction and development. In this study, we have established a reference C. tritonis transcriptome derived from developmental stages (embryo and veliger) and adult tissues. This was used to identify genes associated with cell signalling, such as neuropeptides and G protein-coupled receptors (GPCRs), involved in endocrine and olfactory signalling. A comparison of developmental stages showed that several neuropeptide precursors are exclusively expressed in post-hatch veligers and functional analysis found that FFamide stimulated a significant (20.3%) increase in larval heart rate. GPCRs unique to veligers, and a diversity of rhodopsin-like GPCRs located within adult cephalic tentacles, all represent candidate olfactory receptors. In addition, the cytochrome P450 superfamily, which participates in the biosynthesis and degradation of steroid hormones and lipids, was also found to be expanded with at least 91 genes annotated, mostly in gill tissue. These findings further progress our understanding of C. tritonis with possible application in developing aquaculture methods.

Genetic characterization of ms (3) K81, a paternal effect gene of Drosophila melanogaster.

Genetics ◽  
1995 ◽  
Vol 140 (1) ◽  
pp. 219-229 ◽  
Author(s):  
G K Yasuda ◽  
G Schubiger ◽  
B T Wakimoto
Keyword(s):  
Drosophila Melanogaster ◽  
Developmental Stages ◽  
Normal Function ◽  
Loss Of Function ◽  
Male Sterile ◽  
Specific Product ◽  
Paternal Effect ◽  
Developmental Arrest ◽  
Effect Gene

Abstract The vast majority of known male sterile mutants of Drosophila melanogaster fail to produce mature sperm or mate properly. The ms(3) K81(1) mutation is one of a rare class of male sterile mutations in which sterility is caused by developmental arrest after sperm entry into the egg. Previous studies showed that males homozygous for the K81(1) mutation produce progeny that arrest at either of two developmental stages. Most embryos arrest during early nuclear cycles, whereas the remainder are haploid embryos that arrest at a later stage. This description of the mutant phenotype was based on the analysis of a single allele isolated from a natural population. It was therefore unclear whether this unique paternal effect phenotype reflected the normal function of the gene. The genetic analysis and initial molecular characterization of five new K81 mutations are described here. Hemizygous conditions and heteroallelic combinations of the alleles were associated with male sterility caused by defects in embryogenesis. No other mutant phenotypes were observed. Thus, the K81 gene acted as a strict paternal effect gene. Moreover, the biphasic pattern of developmental arrest was common to all the alleles. These findings strongly suggested that the unusual embryonic phenotype caused by all five new alleles was due to loss of function of the K81+ gene. The K81 gene is therefore the first clear example of a strict paternal effect gene in Drosophila. Based on the embryonic lethal phenotypes, we suggest that the K81+ gene encodes a sperm-specific product that is essential for the male pronucleus to participate in the first few embryonic nuclear divisions.

The transmembrane protein, Tincar, is involved in the development of the compound eye in Drosophila melanogaster

2005 ◽  
Vol 215 (2) ◽  
pp. 90-96 ◽  
Author(s):  
Yuki Hirota ◽  
Kazunobu Sawamoto ◽  
Kuniaki Takahashi ◽  
Ryu Ueda ◽  
Hideyuki Okano
Keyword(s):  
Drosophila Melanogaster ◽  
Compound Eye ◽  
Transmembrane Protein

The 93D heat shock locus of Drosophila melanogaster: modulation by genetic and developmental factors

Genome ◽  
1989 ◽  
Vol 31 (2) ◽  
pp. 677-683 ◽  
Author(s):  
S. C. Lakhotia
Keyword(s):  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Developmental Regulation ◽  
Sequence Divergence ◽  
Unique Region ◽  
Reading Frame ◽  
Beta Alanine ◽  
Functional Conservation ◽  
Duplicate Loci ◽  
93D Locus

The 93D locus in Drosophila melanogaster and the 93D-like loci in other species of Drosophila, collectively termed hsr ω (heat shock RNA omega) locus, display several unique and intriguing features: (i) developmental regulation and selective induction by several agents like benzamide, colchicine, thiamphenicol, vit-B6; (ii) functional conservation in the genus but a very rapid DNA base sequence divergence; (iii) in spite of the rapid DNA sequence divergence, a strong conservation of organization (a 5′ unique region and a 3′ long tandem repeat region) and the pattern of multiple ω transcripts in the genus; (iv) a general nontranslatability of all the three major species of ω transcripts (an ~ 10-kb ω1, a 2.0-kb ω2, and a 1.2-kb ω3 species) although some recent evidence favours translatability of a small open reading frame (~ 23 – 27 amino acid long) in the ω3 transcript; (v) dispensability of the hsr ω locus for heat shock protein synthesis but indispensability for viability of flies. The heat shock inducibility of the 93D locus of D. melanogaster is selectively repressed by (i) combination of heat shock with another inducer of 93D; (ii) rearing of larvae at 10 °C; (iii) heterozygous deficiency for the 93D region; and (iv) conditions that alter levels of beta-alanine. In all cases of repression of the 93D locus during heat shock, the 87A and 87C loci (the two duplicate loci harbouring multiple copies for hsp70 and the alpha–beta repeat sequences (at 87C)) develop unequal puffs. The hsr ω locus appears to be under a complex system of regulation involving autoregulation as well as regulation by other factors in the cell which possibly operate through different control elements on the locus.Key words: benzamide, colchicine, beta-alanine, hsr ω, heat shock puffs, Drosophila.

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