scholarly journals Dual roles for the Drosophila PI 4-kinase Four wheel drive in localizing Rab11 during cytokinesis

2009 ◽  
Vol 187 (6) ◽  
pp. 847-858 ◽  
Author(s):  
Gordon Polevoy ◽  
Ho-Chun Wei ◽  
Raymond Wong ◽  
Zsofia Szentpetery ◽  
Yeun Ju Kim ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Critical Role ◽  
Successful Completion ◽  
Dual Roles ◽  
Recycling Endosome ◽  
Golgi Membranes ◽  
Wheel Drive ◽  
Dividing Cells ◽  
Four Wheel Drive

Successful completion of cytokinesis relies on addition of new membrane, and requires the recycling endosome regulator Rab11, which localizes to the midzone. Despite the critical role of Rab11 in this process, little is known about the formation and composition of Rab11-containing organelles. Here, we identify the phosphatidylinositol (PI) 4-kinase III β Four wheel drive (Fwd) as a key regulator of Rab11 during cytokinesis in Drosophila melanogaster spermatocytes. We show Fwd is required for synthesis of PI 4-phosphate (PI4P) on Golgi membranes and for formation of PI4P-containing secretory organelles that localize to the midzone. Fwd binds and colocalizes with Rab11 on Golgi membranes, and is required for localization of Rab11 in dividing cells. A kinase-dead version of Fwd also binds Rab11 and partially restores cytokinesis to fwd mutant flies. Moreover, activated Rab11 partially suppresses loss of fwd. Our data suggest Fwd plays catalytic and noncatalytic roles in regulating Rab11 during cytokinesis.

The P-element-induced silencing effect of KP transposons is dose dependent in Drosophila melanogaster

Genome ◽  
2011 ◽  
Vol 54 (9) ◽  
pp. 752-762 ◽  
Author(s):  
Alireza Sameny ◽  
John Locke
Keyword(s):  
Drosophila Melanogaster ◽  
Critical Role ◽  
Mutagenic Effect ◽  
P Element ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
P Elements ◽  
Single Well ◽  
Dose Dependent

Transposable elements are found in the genomes of all eukaryotes and play a critical role in altering gene expression and genome organization. In Drosophila melanogaster, transposable P elements are responsible for the phenomenon of hybrid dysgenesis. KP elements, a deletion-derivative of the complete P element, can suppress this mutagenic effect. KP elements can also silence the expression of certain other P-element-mediated transgenes in a process called P-element-dependent silencing (PDS), which is thought to involve the recruitment of heterochromatin proteins. To explore the mechanism of this silencing, we have mobilized KP elements to create a series of strains that contain single, well-defined KP insertions that show PDS. To understand the quantitative role of KP elements in PDS, these single inserts were combined in a series of crosses to obtain genotypes with zero, one, or two KP elements, from which we could examine the effect of KP gene dose. The extent of PDS in these genotypes was shown to be dose dependent in a logarithmic rather than linear fashion. A logarithmic dose dependency is consistent with the KP products interacting with heterochromatic proteins in a concentration-dependent manner such that two molecules are needed to induce gene silencing.

scholarly journals Molecular basis of the dual role of the Mlh1-Mlh3 endonuclease in MMR and in meiotic crossover formation

2021 ◽  
Vol 118 (23) ◽  
pp. e2022704118
Author(s):  
Jingqi Dai ◽  
Aurore Sanchez ◽  
Céline Adam ◽  
Lepakshi Ranjha ◽  
Giordano Reginato ◽  
...  
Keyword(s):  
Meiotic Recombination ◽  
Molecular Basis ◽  
Crystal Packing ◽  
Critical Role ◽  
Holliday Junctions ◽  
Dual Roles ◽  
Terminal Domain ◽  
C Terminus ◽  
Meiotic Crossover

In budding yeast, the MutL homolog heterodimer Mlh1-Mlh3 (MutLγ) plays a central role in the formation of meiotic crossovers. It is also involved in the repair of a subset of mismatches besides the main mismatch repair (MMR) endonuclease Mlh1-Pms1 (MutLα). The heterodimer interface and endonuclease sites of MutLγ and MutLα are located in their C-terminal domain (CTD). The molecular basis of MutLγ’s dual roles in MMR and meiosis is not known. To better understand the specificity of MutLγ, we characterized the crystal structure of Saccharomyces cerevisiae MutLγ(CTD). Although MutLγ(CTD) presents overall similarities with MutLα(CTD), it harbors some rearrangement of the surface surrounding the active site, which indicates altered substrate preference. The last amino acids of Mlh1 participate in the Mlh3 endonuclease site as previously reported for Pms1. We characterized mlh1 alleles and showed a critical role of this Mlh1 extreme C terminus both in MMR and in meiotic recombination. We showed that the MutLγ(CTD) preferentially binds Holliday junctions, contrary to MutLα(CTD). We characterized Mlh3 positions on the N-terminal domain (NTD) and CTD that could contribute to the positioning of the NTD close to the CTD in the context of the full-length MutLγ. Finally, crystal packing revealed an assembly of MutLγ(CTD) molecules in filament structures. Mutation at the corresponding interfaces reduced crossover formation, suggesting that these superstructures may contribute to the oligomer formation proposed for MutLγ. This study defines clear divergent features between the MutL homologs and identifies, at the molecular level, their specialization toward MMR or meiotic recombination functions.

Disruption of selenoprotein biosynthesis affects cell proliferation in the imaginal discs and brain of Drosophila melanogaster

1999 ◽  
Vol 112 (17) ◽  
pp. 2875-2884
Author(s):  
B. Alsina ◽  
M. Corominas ◽  
M.J. Berry ◽  
J. Baguna ◽  
F. Serras
Keyword(s):  
Cell Proliferation ◽  
Drosophila Melanogaster ◽  
Mrna Expression ◽  
Redox Balance ◽  
Imaginal Discs ◽  
Brdu Incorporation ◽  
Pupal Development ◽  
Localization Analysis ◽  
Dividing Cells

The patufet gene encodes the Drosophila melanogaster homologue of selenophosphate synthetase, an enzyme required for selenoprotein synthesis, and appears to have a role in cell proliferation. In this paper we analyse the expression pattern of patufet during the development of imaginal discs and brain as well as the function of this gene in relation to cell proliferation. Wild-type organisms showed a highly dynamic pattern of ptuf mRNA expression during larval and pupal development. Co-localization analysis of ptuf mRNA expression and BrdU incorporation showed high levels of ptuf mRNA in dividing cells and low or undetectable levels in non-dividing cells. In addition, [(75)Se] incorporation revealed a major selenoprotein band of 42 kDa. Mutant organisms showed no selenoprotein synthesis, lower levels of cell proliferation, a higher proportion of cells arrested in G(2) as seen by cyclin B labeling and increased levels of reactive oxygen species (ROS). Because most selenoproteins identified so far are antioxidants, the role of ptuf in cell proliferation through the control of the cellular redox balance is discussed.

A phospholipid kinase regulates actin organization and intercellular bridge formation during germline cytokinesis

Development ◽  
2000 ◽  
Vol 127 (17) ◽  
pp. 3855-3864 ◽  
Author(s):  
J.A. Brill ◽  
G.R. Hime ◽  
M. Scharer-Schuksz ◽  
M.T. Fuller
Keyword(s):  
Critical Role ◽  
Male Meiosis ◽  
Intercellular Bridge ◽  
Bridge Formation ◽  
Actin Organization ◽  
Developmental Context ◽  
Daughter Cells ◽  
Wheel Drive ◽  
Phosphatidylinositol 4 ◽  
Four Wheel Drive

The endgame of cytokinesis can follow one of two pathways depending on developmental context: resolution into separate cells or formation of a stable intercellular bridge. Here we show that the four wheel drive (fwd) gene of Drosophila melanogaster is required for intercellular bridge formation during cytokinesis in male meiosis. In fwd mutant males, contractile rings form and constrict in dividing spermatocytes, but cleavage furrows are unstable and daughter cells fuse together, producing multinucleate spermatids. fwd is shown to encode a phosphatidylinositol 4-kinase (PI 4-kinase), a member of a family of proteins that perform the first step in the synthesis of the key regulatory membrane phospholipid PIP2. Wild-type activity of the fwd PI 4-kinase is required for tyrosine phosphorylation in the cleavage furrow and for normal organization of actin filaments in the constricting contractile ring. Our results suggest a critical role for PI 4-kinases and phosphatidylinositol derivatives during the final stages of cytokinesis.

Role of vision throughout the flight initiation sequence of the Colorado potato beetle

2005 ◽  
Vol 137 (4) ◽  
pp. 420-427
Author(s):  
Gilles Boiteau
Keyword(s):  
Colorado Potato Beetle ◽  
Leptinotarsa Decemlineata ◽  
Critical Role ◽  
Successful Completion ◽  
Flight Chamber ◽  
Potato Beetle ◽  
Key Factor ◽  
Initiation Sequence ◽  
Flight Initiation

AbstractThe role of vision in the flight initiation sequence of the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), was assessed by comparing the behavioral sequences and frequencies of takeoffs of untethered and tethered beetles with and without vision. Results showed that vision plays a critical role in the successful completion of the launch and the maintenance of lift for the adult Colorado potato beetle. The sequence was interrupted immediately before or during the lifting of the mesothoracic legs or before the launch for most blind beetles. Liftoffs of untethered beetles over a 1-h period in a flight chamber were reduced significantly, by 77%, in blind beetles compared with normal beetles. In addition, the preponderance of upward (dispersal) flights following successful liftoffs of normal beetles was replaced by a preponderance of downward (short) flights for blind beetles. Blind beetles that did not have to launch themselves because they were tethered initiated flight as frequently as tethered normal Colorado potato beetles. The duration and repeatability of flights initiated by tethered blind Colorado potato beetles were also similar to those of flights initiated by tethered normal Colorado potato beetles in a stationary environment. Together, the results of the tests with untethered and tethered beetles indicate that vision is a key factor during flight launching and for flight continuation.

scholarly journals CRISP1 as a novel CatSper regulator that modulates sperm motility and orientation during fertilization

2015 ◽  
Vol 210 (7) ◽  
pp. 1213-1224 ◽  
Author(s):  
Juan I. Ernesto ◽  
Mariana Weigel Muñoz ◽  
María A. Battistone ◽  
Gustavo Vasen ◽  
Pablo Martínez-López ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Critical Role ◽  
Cumulus Cells ◽  
Fine Tuning ◽  
Patch Clamping ◽  
Successful Completion ◽  
Tuning Mechanism ◽  
Mammalian Fertilization ◽  
Ca2 Channels

Ca2+-dependent mechanisms are critical for successful completion of fertilization. Here, we demonstrate that CRISP1, a sperm protein involved in mammalian fertilization, is also present in the female gamete and capable of modulating key sperm Ca2+ channels. Specifically, we show that CRISP1 is expressed by the cumulus cells that surround the egg and that fertilization of cumulus–oocyte complexes from CRISP1 knockout females is impaired because of a failure of sperm to penetrate the cumulus. We provide evidence that CRISP1 stimulates sperm orientation by modulating sperm hyperactivation, a vigorous motility required for penetration of the egg vestments. Moreover, patch clamping of sperm revealed that CRISP1 has the ability to regulate CatSper, the principal sperm Ca2+ channel involved in hyperactivation and essential for fertility. Given the critical role of Ca2+ for sperm motility, we propose a novel CRISP1-mediated fine-tuning mechanism to regulate sperm hyperactivation and orientation for successful penetration of the cumulus during fertilization.

scholarly journals The GATOR1 Complex Regulates Metabolic Homeostasis and the Response to Nutrient Stress in Drosophila melanogaster

2016 ◽  
Vol 6 (12) ◽  
pp. 3859-3867 ◽  
Author(s):  
Youheng Wei ◽  
Brad Reveal ◽  
Weili Cai ◽  
Mary A Lilly
Keyword(s):  
Drosophila Melanogaster ◽  
Amino Acid ◽  
Fat Body ◽  
Critical Role ◽  
Nutrient Stress ◽  
Animal Physiology ◽  
Evolutionarily Conserved ◽  
A Cell ◽  
Baseline Levels

Abstract TORC1 regulates metabolism and growth in response to a large array of upstream inputs. The evolutionarily conserved trimeric GATOR1 complex inhibits TORC1 activity in response to amino acid limitation. In humans, the GATOR1 complex has been implicated in a wide array of pathologies including cancer and hereditary forms of epilepsy. However, the precise role of GATOR1 in animal physiology remains largely undefined. Here, we characterize null mutants of the GATOR1 components nprl2, nprl3, and iml1 in Drosophila melanogaster. We demonstrate that all three mutants have inappropriately high baseline levels of TORC1 activity and decreased adult viability. Consistent with increased TORC1 activity, GATOR1 mutants exhibit a cell autonomous increase in cell growth. Notably, escaper nprl2 and nprl3 mutant adults have a profound locomotion defect. In line with a nonautonomous role in the regulation of systemic metabolism, expressing the Nprl3 protein in the fat body, a nutrient storage organ, and hemocytes but not muscles and neurons rescues the motility of nprl3 mutants. Finally, we show that nprl2 and nprl3 mutants fail to activate autophagy in response to amino acid limitation and are extremely sensitive to both amino acid and complete starvation. Thus, in Drosophila, in addition to maintaining baseline levels of TORC1 activity, the GATOR1 complex has retained a critical role in the response to nutrient stress. In summary, the TORC1 inhibitor GATOR1 contributes to multiple aspects of the development and physiology of Drosophila.

scholarly journals Host and Bacterial Factors Control Susceptibility of Drosophila melanogaster to Coxiella burnetii Infection

2017 ◽  
Vol 85 (7) ◽  
Author(s):  
Reginaldo G. Bastos ◽  
Zachary P. Howard ◽  
Aoi Hiroyasu ◽  
Alan G. Goodman
Keyword(s):  
Drosophila Melanogaster ◽  
Coxiella Burnetii ◽  
Q Fever ◽  
Animal Health ◽  
Critical Role ◽  
Content Type ◽  
Imd Pathway ◽  
Tnf Superfamily ◽  
Necrosis Factor

ABSTRACT Coxiella burnetii is the causative agent of Q fever, a zoonotic disease that threatens both human and animal health. Due to the paucity of experimental animal models, little is known about how host factors interface with bacterial components and affect pathogenesis. Here, we used Drosophila melanogaster, in conjunction with the biosafety level 2 (BSL2) Nine Mile phase II (NMII) clone 4 strain of C. burnetii, as a model to investigate host and bacterial components implicated in infection. We demonstrate that adult Drosophila flies are susceptible to C. burnetii NMII infection and that this bacterial strain, which activates the immune deficiency (IMD) pathway, is able to replicate and cause mortality in the animals. We show that in the absence of Eiger, the only known tumor necrosis factor (TNF) superfamily homolog in Drosophila, Coxiella-infected flies exhibit reduced mortality from infection. We also demonstrate that the Coxiella type 4 secretion system (T4SS) is critical for the formation of the Coxiella-containing vacuole and establishment of infection in Drosophila. Altogether, our data reveal that the Drosophila TNF homolog Eiger and the Coxiella T4SS are implicated in the pathogenesis of C. burnetii in flies. The Drosophila/NMII model mimics relevant aspects of the infection in mammals, such as a critical role of host TNF and the bacterial T4SS in pathogenesis. Our work also demonstrates the usefulness of this BSL2 model to investigate both host and Coxiella components implicated in infection.

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