The wingless product is required for cell proliferation in the Malpighian tubule anlage of Drosophila melanogaster

Development ◽  
1992 ◽  
Vol 116 (3) ◽  
pp. 745-754 ◽  
Author(s):  
H. Skaer ◽  
A. Martinez Arias
Keyword(s):  
Cell Proliferation ◽  
Drosophila Melanogaster ◽  
Cell Division ◽  
Malpighian Tubule ◽  
Malpighian Tubules ◽  
Temperature Sensitive ◽  
Over Expression ◽  
Normal Expression ◽  
Segment Polarity ◽  
Sensitive Allele

Cell division in the Malpighian tubules of Drosophila melanogaster depends on the presence of a specialised cell at the tip of each tubule (Skaer, H. le B (1989) Nature 342, 566–569). Here we show that cell division also depends on the normal expression of the segment polarity gene, wingless. The pattern of wingless RNA and protein in developing tubules is consistent with a requirement for wingless for cell division. Analysis of the temporal requirement for wingless using a temperature- sensitive allele confirms that the normal expression of wingless is necessary during cell proliferation in the Malpighian tubules. Over-expression of the gene, induced in a stock containing the wg gene under the control of a heat-shock promoter, results in supernumerary cells in the tubules. We discuss the role of wingless in the cell interactions that govern cell division in the Malpighian tubules.

A Rapid Degradation of Calponin 2 Is Required for Cytokinesis

2021 ◽  
Author(s):  
Airong Qian ◽  
Tzu-Bou Hsieh ◽  
M. Moazzem Hossain ◽  
Jim J.-C. Lin ◽  
J.-P. Jin
Keyword(s):  
Cell Proliferation ◽  
Cell Division ◽  
Underlying Mechanism ◽  
Fluorescent Imaging ◽  
Hek293 Cells ◽  
Computer Assisted ◽  
Over Expression ◽  
Dividing Cells ◽  
Ubiquitin Proteasome ◽  
Dynamic Image Analysis

Calponin 2 is an actin cytoskeleton-associated protein and plays a role in regulating cell motility-related functions such as phagocytosis, migration and division. We previously reported that the expression of calponin 2 inhibits the rate of cell proliferation. To investigate the underlying mechanism, our present study found that the levels of endogenous calponin 2 in NIH3T3 and HEK293 cells rapidly decreased prior to cell division characterized by an absence at the actin contractile ring. In cells lacking endogenous calponin 2, transfective expression of GFP-fusion calponin 2 inhibited cell proliferation similar to that of non-fusion calponin 2. Fluorescent imaging studies of mitotic cells indicated that a proper level of calponin 2 expression and effective degradation during cytokinesis are necessary for normal cell division. Computer-assisted dynamic image analysis of dividing cells revealed that over-expression of calponin 2 significantly affects motile and shape behaviors of cells only on the interval from the start of anaphase to the start of cytokinesis, i.e., the pre-cytokinesis phase, but not on the interval from the start of cytokinesis to 50% completion of cytokinesis. The pre-cytokinesis degradation of calponin 2 was attenuated by MG132 inhibition of the ubiquitin proteasome and inhibitor of protein kinase C (PKC), suggesting that PKC phosphorylation-triggered degradation of calponin 2 could determine the rate of cytokinesis. The novel role of calponin 2 in regulating the rate of cytokinesis may be targeted for therapeutic applications such as in an inhibition of malignant tumor growth.

Studies on the female-sterile phenotype of 1(1)su(f) ts76a, a temperature-sensitive allele of the suppressor of forked mutation in Drosophila melanogaster

Development ◽  
1980 ◽  
Vol 55 (1) ◽  
pp. 247-256
Author(s):  
Thomas G. Wilson
Keyword(s):  
Cell Death ◽  
Drosophila Melanogaster ◽  
Follicle Cell ◽  
Lethal Mutation ◽  
Female Sterility ◽  
Temperature Sensitive ◽  
Egg Chambers ◽  
A Cell ◽  
Sensitive Allele

A new allele of the suppressor of forked [su(f)] mutation in Drosophila melanogaster has been found and designated 1(1)su(f)ts76a. It is temperature-sensitive for suppression of forked (f) and has additional temperature-sensitive phenotypes of lethality, female sterility, and abnormal bristle formation at 29 °C. It closely resembles two other conditional alleles of su(f), 1(1)su(f)ts67g and 1(1)ts726. Female sterility at 29 °C is characterized by both disorganized egg chambers in the ovarioles and also chorion-deficient oocytes. Both of these abnormalities may be the result of premature follicle cell death. The observations on 1(1)su(f)ts76a are consistent with the proposal that the similar allele, 1(1)ts726, is a cell-lethal mutation specifically affecting mitotically active cells.

scholarly journals Loss of DIAPH3, a Formin Family Protein, Leads to Cytokinetic Failure Only under High Temperature Conditions in Mouse FM3A Cells

2020 ◽  
Vol 21 (22) ◽  
pp. 8493
Author(s):  
Hiroki Kazama ◽  
Shu-ichiro Kashiwaba ◽  
Sayaka Ishii ◽  
Keiko Yoshida ◽  
Yuta Yatsuo ◽  
...  
Keyword(s):  
Cell Division ◽  
High Temperature ◽  
Temperature Sensitive ◽  
Dependent Manner ◽  
Temperature Dependent ◽  
Over Expression ◽  
Temperature Conditions ◽  
Family Protein ◽  
Temperature Environment ◽  
Ts Mutant

Cell division is essential for the maintenance of life and involves chromosome segregation and subsequent cytokinesis. The processes are tightly regulated at both the spatial and temporal level by various genes, and failures in this regulation are associated with oncogenesis. Here, we investigated the gene responsible for defects in cell division by using murine temperature-sensitive (ts) mutant strains, tsFT101 and tsFT50 cells. The ts mutants normally grow in a low temperature environment (32 °C) but fail to divide in a high temperature environment (39 °C). Exome sequencing and over-expression analyses identified Diaph3, a member of the formin family, as the cause of the temperature sensitivity observed in tsFT101 and tsFT50 cells. Interestingly, Diaph3 knockout cells showed abnormality in cytokinesis at 39 °C, and the phenotype was rescued by re-expression of Diaph3 WT, but not Diaph1 and Diaph2, other members of the formin family. Furthermore, Diaph3 knockout cells cultured at 39 °C showed a significant increase in the level of acetylated α-tubulin, an index of stabilized microtubules, and the level was reduced by Diaph3 expression. These results suggest that Diaph3 is required for cytokinesis only under high temperature conditions. Therefore, our study provides a new insight into the mechanisms by which regulatory factors of cell division function in a temperature-dependent manner.

Long noncoding RNA LINC00968 inhibits proliferation, migration and invasion of lung adenocarcinoma through  targeting  miR-22-5p / CDC14A  axis

2020 ◽  
Author(s):  
Chao Wu ◽  
Xuzhao Bian ◽  
Liyuan Zhang ◽  
Yang Wu ◽  
Tianli Pei ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Division ◽  
Lung Adenocarcinoma ◽  
Cell Lines ◽  
Cell Division Cycle ◽  
Cell Counting ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Over Expression ◽  
Node Metastasis

Abstract Background Lung adenocarcinoma (LUAD) is a high aggressive human cancer which usually diagnosed at advanced stages. Accumulating evidences indicate that long noncoding RNAs (lncRNAs) are crucial participants in LUAD progression. Methods The mRNA levels of LINC00968, miR-22-5p and cell division cycle 14A (CDC14A) were measured using quantitative real-time PCR. Cell proliferation was evaluated using cell counting kit-8 and flow cytometry. Cell migration and cell invasion were assessed by wound healing and transwell assay, respectively. The interactions between LINC00968 and miR-22-5p were validated by RNA immunoprecipitation, RNA pull down and luciferase reporter assay. Results We found that lncRNA LINC00968 was significantly down-regulated in LUAD tissues and cell lines. LINC00968 level was positively correlated to survival rate, and negatively correlated to tumor node metastasis stage, tumor size and lymph node metastasis of LUAD patients. LINC00968 over-expression in LUAD cells inhibited cell proliferation and induced cell cycle arrest at G1 phase. LINC00968 over-expression also suppressed migration, invasion and epithelial mesenchymal transition (EMT) as evidenced by elevated E-cadherin, decreased N-cadherin, TWIST and SNAIL levels. We further validated that LINC00968 localized in cytoplasma and acted as an upstream of microRNA miR-22-5p, which was up-regulated in LUAD tissues and cell lines. Besides, elevated miR-22-5p expression abolished the effect of LINC00968 over-expression on LUAD progression including in vivo tumor growth. In addition, we first validated that cell division cycle 14A (CDC14A), which was down-regulated in LUAD tissues, was a downstream target of miR-22-5p. We over-expressed CDC14A in LUAD cells and miR-22-5p induced LUAD progression was partially reversed. Conclusion our study demonstrated that LINC00968 inhibited proliferation, migration and invasion of LUAD by sponging miR-22-5p and further restoring CDC14A. This novel regulatory network might provide us with promising diagnostic and therapeutic target in LUAD treatment.

Molecular characterization of the Drosophila melanogaster urate oxidase gene, an ecdysone-repressible gene expressed only in the malpighian tubules

1990 ◽  
Vol 10 (10) ◽  
pp. 5114-5127
Author(s):  
L L Wallrath ◽  
J B Burnett ◽  
T B Friedman
Keyword(s):  
Drosophila Melanogaster ◽  
Amino Acid ◽  
Regulatory Element ◽  
Malpighian Tubule ◽  
Amino Acid Sequences ◽  
Malpighian Tubules ◽  
P Element ◽  
Urate Oxidase ◽  
Base Pairs ◽  
Oxidase Gene

The urate oxidase (UO) gene of Drosophila melanogaster is expressed during the third-instar larval and adult stages, exclusively within a subset of cells of the Malpighian tubules. The UO gene contains a 69-base-pair intron and encodes mature mRNAs of 1,224, 1,227, and 1,244 nucleotides, depending on the site of 3' endonucleolytic cleavage prior to polyadenylation. A direct repeat, 5'-AAGTGAGAGTGAT-3', is the proposed cis-regulatory element involved in 20-hydroxyecdysone repression of the UO gene. The deduced amino acid sequences of UO of D. melanogaster, rat, mouse, and pig and uricase II of soybean show 32 to 38% identity, with 22% of amino acid residues identical in all species. With use of P-element-mediated germ line transformation, 826 base pairs 5' and approximately 1,200 base pairs 3' of the D. melanogaster UO transcribed region contain all of the cis elements allowing for appropriate temporal regulation and Malpighian tubule-specific expression of the UO gene.

scholarly journals Zinc storage granules in the Malpighian tubules of Drosophila melanogaster

2017 ◽  
Author(s):  
Carlos Tejeda-Guzmán ◽  
Abraham Rosas-Arellano ◽  
Thomas Kroll ◽  
Samuel M. Webb ◽  
Martha Barajas-Aceves ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Malpighian Tubule ◽  
Adaptor Protein ◽  
Mossy Fibers ◽  
Membrane Transporters ◽  
Malpighian Tubules ◽  
Pancreatic Cells ◽  
Storage Granules ◽  
Hermansky Pudlak Syndrome ◽  
Zinc Storage

ABSTRACTMembrane transporters and sequestration mechanisms concentrate metal ions differentially into discrete subcellular microenvironments for usage in protein cofactors, signaling, storage, or excretion. Here we identify zinc storage granules as the insect’s major zinc reservoir in primary Malpighian tubule epithelial cells of Drosophila melanogaster. The concerted action of Adaptor Protein-3, Rab32, HOPS and BLOC complexes as well as of the white-scarlet (ABCG2-like) and ZnT35C transporters is required for zinc storage granule biogenesis. Due to similar lysosome related organelle defects, patients with Hermansky-Pudlak syndrome may lack zinc granules in beta pancreatic cells, intestinal paneth cells and presynaptic vesicles of hippocampal mossy fibers.

Characterization of theDrosophila melanogasteralkali-metal/proton exchanger (NHE) gene family

2001 ◽  
Vol 204 (21) ◽  
pp. 3703-3716 ◽  
Author(s):  
Maria E. Giannakou ◽  
Julian A. T. Dow
Keyword(s):  
Drosophila Melanogaster ◽  
Genomic Sequence ◽  
Malpighian Tubule ◽  
Malpighian Tubules ◽  
Na Channel ◽  
Rt Pcr ◽  
Multiple Transcripts ◽  
Homology Searching ◽  
Amiloride Derivatives

SUMMARYThe NHE family of Na+/H+ exchangers is believed to play an essential role in animals, but may play an additional, specialised epithelial role in insects. The pharmacological sensitivity of the Drosophila melanogaster Malpighian tubule to a range of amiloride derivatives was shown to be consistent with an effect on an exchanger, rather than a Na+ channel. Consistent with this, no degenerin/epithelial Na+ channel (ENaC) genes could be detected in Malpighian tubules by reverse transcriptase/polymerase chain reaction (RT-PCR). Using a low-stringency homology searching, three members of the NHE family were identified in the genomic sequence of Drosophila melanogaster, although only two genes were represented as expressed sequence tags. All three genes (DmNHE1 at cytological position 21B1, DmNHE2 at 39B1 and DmNHE3 at 27A1) were found by RT-PCR to be widely expressed, and one (DmNHE2) was shown to have multiple transcripts. The putative translations of the three genes mark them as distantly related members of the family, inviting the possibility that they may serve distinct roles in insects.

Genetic analysis of the Drosophila cdc2 homolog

Development ◽  
1993 ◽  
Vol 117 (1) ◽  
pp. 219-232 ◽  
Author(s):  
B. Stern ◽  
G. Ried ◽  
N.J. Clegg ◽  
T.A. Grigliatti ◽  
C.F. Lehner
Keyword(s):  
Cell Proliferation ◽  
Sequence Analysis ◽  
Genetic Analysis ◽  
S Phase ◽  
P Element ◽  
Temperature Sensitive ◽  
Cdc2 Gene ◽  
G2 Phase ◽  
Sensitive Allele ◽  
Mitotic Proliferation

We have identified mutations in the Drosophila cdc2 gene. The recessive lethality of these mutant alleles was rescued after P-element-mediated transformation with a genomic cdc2 fragment. Sequence analysis of amorphic alleles revealed non-conservative exchanges in evolutionary conserved positions. These alleles caused lethality at the larval-pupal interphase due to the absence of imaginal tissues. Embryonic lethality resulted when the maternal Dm cdc2 contribution was reduced through the use of a temperature-sensitive allele. Dm cdc2 function, therefore, is essential for cell proliferation throughout development. Dm cdc2 function is clearly required for mitosis, but no evidence for a requirement in S-phase was obtained. The reversible block of the mitotic proliferation which was observed in the PNS of mutant embryos occurred exclusively in the G2-phase. Moreover, while the mitotic proliferation of imaginal cells was blocked in the amorphic mutant larvae, non-imaginal larval cells continued to grow and endoreplicate their DNA. The Dm cdc2 mutant phenotype could neither be rescued with Dm cdc2c (encoding a cdc2-like kinase) nor enhanced by a reduction of the Dm cdc2c gene dose. These results indicate that the Dm cdc2- and Dm cdc2c-kinases control different processes.

Organ growth without cell division: somatic polyploidy in a moth, Ephestia kuehniella

Genome ◽  
2012 ◽  
Vol 55 (11) ◽  
pp. 755-763 ◽  
Author(s):  
Lydia Buntrock ◽  
František Marec ◽  
Sarah Krueger ◽  
Walther Traut
Keyword(s):  
Cell Division ◽  
Cell Growth ◽  
Larval Instar ◽  
Malpighian Tubule ◽  
Silk Gland ◽  
Ephestia Kuehniella ◽  
Malpighian Tubules ◽  
Organ Growth ◽  
Gland Cells ◽  
First Instar

Organ growth depends on cell division and (or) cell growth. Here, we present a study on two organs whose growth depends entirely on cell growth, once they are formed in the embryo: Malpighian tubules and silk glands of the flour moth, Ephestia kuehniella . Between first and last larval instar, the volume of Malpighian tubule cells increases by a factor of ∼1800 and that of silk gland cells by a factor of ∼3100. We determined the number of endocyles required to reach these stages by Feulgen cytometry. Cells of Malpighian tubules were in the 2C stage in first instar larvae and reached 1024C after 9 endocycles in last instar larvae (1C = 0.45 pg DNA). Silk gland cells already reached a DNA content of 8C–16C in first instar larvae and attained up to 8192C in last instar larvae after a total of 12 endocycles. The nuclei were small and more or less spherical in first instar larvae, but they were huge, flat, and bizarrely branched in last instar larvae. We consider branching as a compensatory adaptation to improve molecular traffic between nucleus and cytoplasm in these excessively large and highly polyploid cells (i) by reducing the mean distance between nucleus and cytoplasm and (ii) by enlarging the surface-to-volume ratio of these nuclei.

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