Kendrin/pericentrin-B, a centrosome protein with homology to pericentrin that complexes with PCM-1

2001 ◽  
Vol 114 (4) ◽  
pp. 797-809 ◽  
Author(s):  
Q. Li ◽  
D. Hansen ◽  
A. Killilea ◽  
H.C. Joshi ◽  
R.E. Palazzo ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Evolutionary Relationship ◽  
Coiled Coil ◽  
Maturation Process ◽  
Microtubule Depolymerization ◽  
Microtubule Nucleation ◽  
Sequence Analyses ◽  
Late Event ◽  
Functional Complex ◽  
Centrosome Protein

The centrosome is responsible for nucleating microtubules and performing other cellular roles. To define the organization of the centrosome more completely, a human anti-centrosome serum was used to screen a human cDNA library, and a cDNA encoding a >350 kDa centrosome protein was identified. Sequence analyses revealed that this novel centrosome protein contains two coiled-coil domains bounded by non-coiled regions. The N-terminal region of the protein, named pericentrin-B, shares 61% identity (75% similarity) with pericentrin, suggesting an evolutionary relationship between these proteins. Antibodies against pericentrin-B stain centrosomes at all stages of the cell cycle, and pericentrin-B remains associated with centrosomes following microtubule depolymerization. Immunodepletion of neither pericentrin-B nor PCM-1 from cellular extracts inhibited the ability of salt-stripped centrosomes to recover microtubule nucleation potential, demonstrating that neither protein plays a key role in microtubule nucleation processes. Moreover, the binding of both PCM-1 and pericentrin-B with salt-stripped centrosomes required intact microtubules, demonstrating that the association of PCM-1 and pericentrin-B with centrosomes is a late event in the centrosome maturation process. Finally, pericentrin-B and PCM-1 coimmunoprecipitate, suggesting that PCM-1 and pericentrin-B form a functional complex in cells. This observation may help to explain the generation of anti-centrosome autoantibodies in certain autoimmune patients and may be important for centrosome function.

scholarly journals Characterization of a Drosophila Centrosome Protein CP309 That Shares Homology with Kendrin and CG-NAP

2004 ◽  
Vol 15 (1) ◽  
pp. 37-45 ◽  
Author(s):  
Shin-ichi Kawaguchi ◽  
Yixian Zheng
Keyword(s):  
Cell Cycle ◽  
Drosophila Melanogaster ◽  
Coiled Coil ◽  
Animal Cells ◽  
Microtubule Nucleation ◽  
Small Complex ◽  
Biochemical Studies ◽  
Ring Complex ◽  
Centrosome Protein

The centrosome in animal cells provides a major microtubule-nucleating site that regulates the microtubule cytoskeleton temporally and spatially throughout the cell cycle. We report the identification in Drosophila melanogaster of a large coiled-coil centrosome protein that can bind to calmodulin. Biochemical studies reveal that this novel Drosophila centrosome protein, centrosome protein of 309 kDa (CP309), cofractionates with the γ-tubulin ring complex and the centrosome-complementing activity. We show that CP309 is required for microtubule nucleation mediated by centrosomes and that it interacts with the γ-tubulin small complex. These findings suggest that the microtubule-nucleating activity of the centrosome requires the function of CP309.

scholarly journals Human Enhancer of Invasion-Cluster, a Coiled-Coil Protein Required for Passage through Mitosis

2004 ◽  
Vol 24 (9) ◽  
pp. 3957-3971 ◽  
Author(s):  
Margret B. Einarson ◽  
Edna Cukierman ◽  
Duane A. Compton ◽  
Erica A. Golemis
Keyword(s):  
Cell Cycle ◽  
Coiled Coil ◽  
Cell Cycle Checkpoints ◽  
Mitotic Spindles ◽  
Human Genes ◽  
Hydroxyurea Treatment ◽  
Evolutionarily Conserved ◽  
Defects Analysis ◽  
Spindle Function

ABSTRACT In a cross-species overexpression approach, we used the pseudohyphal transition of Saccharomyces cerevisiae as a model screening system to identify human genes that regulate cell morphology and the cell cycle. Human enhancer of invasion-cluster (HEI-C), encoding a novel evolutionarily conserved coiled-coil protein, was isolated in a screen for human genes that induce agar invasion in S. cerevisiae. In human cells, HEI-C is primarily localized to the spindle during mitosis. Depletion of HEI-C in vivo with short interfering RNAs results in severe mitotic defects. Analysis by immunofluorescence, flow cytometry analysis, and videomicroscopy indicates that HEI-C-depleted cells form metaphase plates with normal timing after G2/M transition, although in many cases cells have disorganized mitotic spindles. Subsequently, severe defects occur at the metaphase-anaphase transition, characterized by a significant delay at this stage or, more commonly, cellular disintegration accompanied by the display of classic biochemical markers of apoptosis. These mitotic defects occur in spite of the fact that HEI-C-depleted cells retain functional cell cycle checkpoints, as these cells arrest normally following nocodazole or hydroxyurea treatment. These results place HEI-C as a novel regulator of spindle function and integrity during the metaphase-anaphase transition.

scholarly journals Characterization of the Crithidia fasciculata mRNA Cycling Sequence Binding Proteins

2001 ◽  
Vol 21 (14) ◽  
pp. 4453-4459 ◽  
Author(s):  
Riaz Mahmood ◽  
Bidyottam Mittra ◽  
Jane C. Hines ◽  
Dan S. Ray
Keyword(s):  
Cell Cycle ◽  
Protein Complexes ◽  
Coiled Coil ◽  
High Specificity ◽  
Binding Activity ◽  
Mrna Levels ◽  
Crithidia Fasciculata ◽  
Cysteine Motif ◽  
Cell Extracts ◽  
Sequence Elements

ABSTRACT The Crithidia fasciculata cycling sequence binding protein (CSBP) binds with high specificity to sequence elements in several mRNAs that accumulate periodically during the cell cycle. Mutations in these sequence elements abolish both cycling of the mRNA and binding of CSBP. Two genes, CSBPA andCSBPB, encoding putative subunits of CSBP have been cloned and were found to be present in tandem on the same DNA molecule and to be closely related. CSBPA andCSBPB are predicted to encode proteins with sizes of 35.6 and 42.0 kDa, respectively. Both CSBPA and CSBPB proteins have a predicted coiled-coil domain near the N terminus and a novel histidine and cysteine motif near the C terminus. The latter motif is conserved in other trypanosomatid species. Gel sieving chromatography and glycerol gradient sedimentation results indicate that CSBP has a molecular mass in excess of 200 kDa and an extended structure. Recombinant CSBPA and CSBPB also bind specifically to the cycling sequence and together can be reconstituted to give an RNA gel shift similar to that of purified CSBP. Proteins in cell extracts bind to an RNA probe containing six copies of the cycling sequence. The RNA-protein complexes contain both CSBPA and CSBPB, and the binding activity cycles in near synchrony with target mRNA levels.CSBPA and CSBPB mRNA and protein levels show little variation throughout the cell cycle, suggesting that additional factors are involved in the cyclic binding to the cycling sequence elements.

scholarly journals The cell cycle state defines TACC3 as a regulator gene in glioblastoma

2020 ◽  
Author(s):  
Holly Briggs ◽  
Euan S. Polson ◽  
Bronwyn K. Irving ◽  
Alexandre Zougman ◽  
Ryan K. Mathew ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Gene Networks ◽  
Coiled Coil ◽  
Cell Lineage ◽  
Progression Free Survival ◽  
Low Grade ◽  
Loss Of Function ◽  
Feature Maps ◽  
Clinical Databases ◽  
Lineage Specificity

AbstractOverexpression and mitosis-promoting roles of Transforming acidic coiled-coil containing protein 3 (TACC3) are well-established in many cancers, including glioblastoma (GBM). However, the effector gene networks downstream of TACC3 remain poorly defined, partly due to an incomplete understanding of TACC3 cell lineage specificity and its dynamic role during the cell cycle. Here, we use a patient-derived GBM model to report that TACC3 predominantly resides in the GBM cell cytoplasm, while engaging in gene regulation temporally as defined by the cell cycle state. TACC3 loss-of-function, cell cycle stage-specific transcriptomics, and unsupervised self-organizing feature maps revealed pathways (including Hedgehog signalling) and individual genes (including HOTAIR) that exhibited anticorrelated expression phenotypes across interphase and mitosis. Furthermore, this approach identified a set of 22 TACC3-dependent transcripts in publicly-available clinical databases that predicted poor overall and progression-free survival in 162 GBM and 514 low-grade glioma patient samples. These findings uncover TACC3-dependent genes as a function of TACC3 cell cycle oscillation, which is important for TACC3-targeting strategies, and for predicting poor outcomes in brain cancer patients.

scholarly journals Potential Common Key Genes Associated with Chronic Periodontitis and Low Birth Weight: A Case Control Study Using Bioinformatics Analysis of Pooled mRNA Expression Datasets

2020 ◽  
Author(s):  
Asami Suzuki ◽  
Tetsuro Horie ◽  
Akihito Nakai ◽  
Eriko Kikuchi ◽  
Yukihiro Numabe
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Birth Weight ◽  
Low Birth Weight ◽  
Chronic Periodontitis ◽  
Web Application ◽  
Coiled Coil ◽  
Growth Development ◽  
Key Genes ◽  
Upstream Regulators

Abstract Background: Chronic periodontitis (CP) is a multifactorial disease associated with many systemic diseases. However, the precise association between CP and low birth weight (LBW) remains unclear. Therefore, this study aimed to elucidate common differentially expressed genes (DEGs), biomarker candidates, and upstream regulators related to key genes between CP and LBW.Methods: We investigated molecular relations and biomarker candidates using pooled microarray datasets of CP (GSE12484) and LBW (GSE29807) in the Gene Expression Omnibus (GEO). Datasets were analyzed for common DEGs using GEO2R, an R-based web application for GEO data analysis. Common DEGs, biomarker candidates, and upstream regulators in DEGs between CP and LBW were analyzed using the Database for Annotation Visualization and Integrated Discovery (DAVID), Search Tool for the Retrieval of Interacting Genes (STRING), and QIAGEN’s Ingenuity Pathway Analysis (IPA).Results: Three significantly upregulated and 20 significantly downregulated common DEGs between CP and LBW were identified. Some biological processes and pathways of these downregulated genes were associated with the cell cycle. Biomarker candidates among common DEGs were proline-rich coiled-coil 2A (PPRC2A), topoisomerase (DNA) II alpha (TOP2A), neural cell adhesion molecule 1 (NCAM1), and calcium channel, voltage-dependent, alpha 2/delta subunit 3 (CACNA2D3). Many upstream regulators of these biomarker candidates were factors associated with inflammation, immunity, the cell cycle, and growth development, and were hormones related to pregnancy.Conclusions: The results of this study suggest that PPRC2A, TOP2A, NCAM1, and CACNA2D3 are common biomedical key genes between CP and LBW. The expression states of these genes, which are related to inflammation, hormones, the cell cycle, and growth development, were common in both CP and LBW in blood. To the best of our knowledge, the relations of PPRC2A, TOP2A, and CACNA2D3 to CP and LBW are reported for the first time. Thus, in the bloodstream, inflammatory-related upstream regulators of these key genes may control gene expression associated with fetal growth, and conversely, changes in female hormones due to pregnancy may affect the progress of CP.

scholarly journals The Novel Nature Microtubule Inhibitor Ivalin Induces G2/M Arrest and Apoptosis in Human Hepatocellular Carcinoma SMMC-7721 Cells In Vitro

Medicina ◽  
2019 ◽  
Vol 55 (8) ◽  
pp. 470 ◽  
Author(s):  
Fangyuan Liu ◽  
Shiqi Lin ◽  
Caiyun Zhang ◽  
Jiahui Ma ◽  
Zhuo Han ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Cycle ◽  
Cell Cycle Arrest ◽  
Western Blotting ◽  
Network Formation ◽  
Cyclin B1 ◽  
Human Hepatocellular Carcinoma ◽  
Microtubule Depolymerization ◽  
Microtubule Network ◽  
Cycle Arrest

Background and Objectives: Microtubules are an attractive target for cancer chemotherapy. Previously, we reported that Ivalin exhibited excellent anti-migration and anti-invasion activities in human breast cancer cells. Here, we examined the microtubule inhibition effect of Ivalin in human hepatocellular carcinoma SMMC-7721 cells. Materials and Methods: We used the 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) assay to evaluate the cell proliferation effect of Ivalin and flow cytometry analysis to detect the apoptotic and cell cycle arrest effects of Ivalin. Immunofluorescence staining was used to measure the effect of Ivalin on the cytoskeleton network, and Western blotting was used to detect the expression levels of Bax, Bcl-2, Cdc2, phosphor-Cdc2, Cdc25A, Cyclin B1, and tubulin. Results: Ivalin induced cell cycle G2/M arrest and subsequent triggered apoptosis in human hepatocellular carcinoma SMMC-7721 cells. Furthermore, microtubules were shown to be involved in Ivalin-meditated apoptosis. In this connection, Ivalin treatment suppressed cellular microtubule network formation by regulating microtubule depolymerization. Moreover, Western blotting revealed Cdc25A and Cyclin B1 were upregulated in Ivalin-meditated cell cycle arrest. Subsequently, the induction of Bax (a proapoptotic protein) and reduction of Bcl-2 (an anti-apoptotic protein) expression were observed in Ivalin-treated SMMC-7721 cells. Conclusion: Ivalin induced microtubule depolymerization, then blocked cells in mitotic phase, and eventually resulted in apoptosis in SMMC-7721 cells. Collectively, these data indicate that Ivalin, acting as a novel inhibitor of microtubules, could be considered as a promising lead in anticancer drug development.

Rho-associated coiled-coil kinase (ROCK) regulates cell cycle of SH-SY5Y cell through β-catenin/TCF pathway

2007 ◽  
Vol 58 ◽  
pp. S208
Author(s):  
Shuken Boku ◽  
Shin Nakagawa ◽  
Katsuji Suzuki ◽  
Takuya Masui ◽  
Mai Kihara ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Coiled Coil

scholarly journals Centrosome maturation: Measurement of microtubule nucleation throughout the cell cycle by using GFP-tagged EB1

2004 ◽  
Vol 101 (6) ◽  
pp. 1584-1588 ◽  
Author(s):  
M. Piehl ◽  
U. S. Tulu ◽  
P. Wadsworth ◽  
L. Cassimeris
Keyword(s):  
Cell Cycle ◽  
Microtubule Nucleation
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