• Title/Summary/Keyword: spindle assembly

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Mad1p, a Component of the Spindle Assembly Checkpoint in Fission Yeast, Suppresses a Novel Septation-defective Mutant, sun1, in a Cell Division Cycle

  • Kim In G.;Rhee Dong K.;Jeong Jae W.;Kim Seong C.;Won Mi S.;Song Ki W.;Kim Hyong B.
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2002.10a
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    • pp.162-172
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    • 2002
  • Schizosaccharomyces pombe is suited for the study of cytokinesis as it divides by forming a septum in the middle of the cell at the end of mitosis. To enhance our understanding of the cytokinesis, we have carried out a genetic screen for temperature-sensitive S. pombe mutants that show defects in septum formation and cell division. Here we present the isolation and characterization of a new temperature-sensitive mutant, sun1(septum uncontrolled), which undergoes uncontrolled septation during cell division cycle at restrictive temperature $(37^{\circ}C)$. In sun1 mutant, actin ring and septum are positioned at random locations and angles, and nuclear division cycle continues. These observations suggest that the sun] gene product is required for the proper placement of the actin ring as well as precise septation. The sun] mutant is monogenic recessive mutation unlinked to previously known various cdc genes of S. pombe. In a screen for $sunl^+$ gene to complement the sun] mutant, we have cloned a gene, $susl^+$(suppressor of sun1 mutant), that encodes a protein of 689 amino acids. The predicted amino acid sequence of $susl^+$ gene is similar to the human hMadlp and Saccharomyces cerevisiae Mad1p, a component of the spindle checkpoint in eukaryotic cells. The null mutant of $susl^+$ gene grows normally at various temperatures and has the increased sensitivity to anti-microtubule drug, while $susl^+$ mutant shows no sensitivity to microtubule destabilizing drugs. The putative S. pombe Sus1p directly interacts with S. pombe Mad2p in yeast two-hybrid assays. These data suggest that the newly isolated susr gene encodes S. pombe Mad1p and suppresses sun] mutant defective in controlled septation in a cell division cycle.

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Microtubule and Chromatin Organization in Bovine Oocytes following Intracytoplasmic Injection of Spermatozoon, Sperm Head and Tail (소 난자에 있어서 세포질내 정자, 정자두부.미부 주입 후 미세소관과 염색질의 구조변화)

  • Do, J.T.;Jun, S.H.;Choi, J.T.;Kang, Y.S.;Lee, B.Y.;Kim, S.B.;Kim, N.H.;Lee, H.T.;Chung, K.S.
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.3
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    • pp.301-309
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    • 1997
  • The objective of this study was to determine the microtubule assembly and chromatin configuration during the first cell cycle in bovine oocytes following injection of spermatozoon, sperm head and tail. The microtubule and chromatin configuration was imaged with fluorescent labeled monoclonal ${\alpha}$-tubulin antibody and propidium iodide under laser scanning confocal microscope. Microtubule and chromatin dynamics in bovine oocytes following intracytoplasmic sperm injection (ICSI) were not different from those observed during in vitro fertilization (IVF). Following ICSI, the microtubular aster was observed around sperm midpiece. During pronuclear formation, the sperm aster was enlarged and seen around male and female pronuclei. At mitotic metaphase, the microtubular spindle assemble astral poles and chromosomes were aligned on the spindle equator. At mitosis, asters were concentrated to each spindle pole and they filled the cytoplasm. After injection of the isolated sperm head, the microtubular aster was not seen around sperm head in any cases (0/18). Instead, microtubules were organized from the cytoplasm, which filled the whole cytoplasm during pronuclear apposition. These microtubules seem to move male and female pronuclei. These results suggest that isolated sperm head can develop into normal pronucleus in mature bovine oocytes, and competent to participate syngamy with the ootid chromatin. The functional microtubules following isolated sperm head injection in bovine oocytes appeared to be organized solely from maternal stores.

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Rotordynamic Characteristics Analysis of Turbocharger Turbine for Spin Test (터보차져용 터빈의 스핀 테스트를 위한 로터다이나믹 특성분석)

  • Kim, Byung-Ok;Yang, Sung-Jin;Lee, Myung-Ho
    • The KSFM Journal of Fluid Machinery
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    • v.14 no.6
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    • pp.91-95
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    • 2011
  • This paper deals with the detail rotordynamic analysis for the assembly rotor composed of turbine wheel, turbine shaft, connecting arbor, and flange & spindle in order to perform the spin test of turbocharger turbine. Prior to rotordynamic analysis, the 1st spin test was performed but the test was failed by excess vibration in the neighborhood rated speed. It is the reason for this fail that the separation margin between the rated speed and critical speed is not enough, confirmed by rotordynamic analysis results. Since then, the dimension of turbine shaft was modified and the critical speeds were again reviewed for modified assmebly rotor. In results, the separation margin between the rated speed and critical speed is over 20% and then the 2nd spin test was performed successfully. In preparing spin test for turbine, compressor wheels and etc., the geometry design of connecting arbor and dimension of rough machining should be reviewed by considering rotordynamic results, and the separation margin should be enough for successful spin test.

Polo-Like Kinases (Plks), a Key Regulator of Cell Cycle and New Potential Target for Cancer Therapy

  • Lee, Su-Yeon;Jang, Chuljoon;Lee, Kyung-Ah
    • Development and Reproduction
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    • v.18 no.1
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    • pp.65-71
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    • 2014
  • Cell cycle process is regulated by a number of protein kinases and among them, serine/threonine kinases carry phosphate group from ATP to substrates. The most important three kinase families are Cyclin-dependent kinase (Cdk), Polo-like kinase (Plk), and Aurora kinase. Polo-like kinase family consists of 5 members (Plk1-Plk5) and they are involved in multiple functions in eukaryotic cell division. It regulates a variety of aspects such as, centrosome maturation, checkpoint recovery, spindle assembly, cytokinesis, apoptosis and many other features. Recently, it has been reported that Plks are related to tumor development and over-expressed in many kinds of tumor cells. When injected the anti-Plk antibody into human cells, the cells show aneuploidy, and if inhibit Plks, most of the mitotic cell division does not proceed properly. For that reasons, many inhibitors of Plk have been recently emerged as new target for remedy of the cancer therapeutic research. In this paper, we reviewed briefly the characteristics of Plk families and how Plks work in regulating cell cycles and cancer formation, and the possibilities of Plks as target for cancer therapy.

Centrobin/Nip2 Expression In Vivo Suggests Its Involvement in Cell Proliferation

  • Lee, Jungmin;Kim, Sunmi;Jeong, Yeontae;Rhee, Kunsoo
    • Molecules and Cells
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    • v.28 no.1
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    • pp.31-36
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    • 2009
  • Centrobin/Nip2 was initially identified as a centrosome protein that is critical for centrosome duplication and spindle assembly. In the present study, we determined the expression and subcellular localization of centrobin in selected mouse tissues. Immunoblot analysis revealed that the centrobin-specific band of 100 kDa was detected in all tissues tested but most abundantly in the thymus, spleen and testis. In the testis, centrobin was localized at the centrosomes of spermatocytes and early round spermatids, but no specific signal was detected in late round spermatids and elongated spermatids. Our results also revealed that the centrosome duplication occurs at interphase of the second meiotic division of the mouse male germ cells. The centrobin protein was more abundant in the mitotically active ovarian follicular cells and thymic cortex cells than in non-proliferating corpus luteal cells and thymic medullary cells. The expression pattern of centrobin suggests that the biological functions of centrobin are related to cell proliferation. Consistent with the proposal, we observed reduction of the centrobin levels when NIH3T3 became quiescent in the serum-starved culture conditions. However, a residual amount of centrobin was also detected at the centrosomes of the resting cells, suggesting its role for maintaining integrity of the centrosome, especially of the daughter centriole in the cells.

Fate of Donor Centrosome and Microtubule Dynamics of Porcine Somatic Cell Nuclear Transfer Embryos

  • Kwon, Dae-Jin;Park, Choon-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Reproductive and Developmental Biology
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    • v.34 no.2
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    • pp.73-79
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    • 2010
  • We investigated the microtubule dynamics, including the inheritance of donor centrosomes and the mitotic spindle assembly occurring during the first mitosis of somatic cell nuclear transfer (SCNT) embryos in pigs. SCNT embryos were fixed 15 min and 1 h after fusion in order to assess the inheritance pattern of the donor centrosome. The distribution and dynamic of the centrosome and microtubule during the first mitotic phase of SCNT embryos were also evaluated. The frequency of embryos evidencing $\gamma$-tubulin spots (centrosome) was 93.2% in the SCNT embryos 15 min after fusion. In the majority of the SCNT embryos (61.5%), however, no centrosome was observed 1 h after fusion. The frequency of the embryos with no or abnormal mitotic spindles 20 h after fusion was 19.6%. The $\gamma$-tubulin spots were detected near the nuclei of somatic cells regardless of cell cycle phase, whereas $\gamma$-tubulin spots in the SCNT embryos were observed only during the inter-anaphase transition. These results showed that the donor centrosome is inherited into the SCNT embryos, but failed to assemble the normal mitotic spindles during first mitotic phase in some SCNT embryos.

Effect of Surface Roughness on Cutting Conditions in CNC lathe C-Axis Milling Arc Cutting (CNC선반 C축 밀링 원호가공에서 절삭조건이 표면 거칠기에 미치는 영향)

  • Shin, Kuk-Sik
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.13 no.4
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    • pp.99-105
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    • 2014
  • The domestic airline industry undertakes the production of finished products by assembling existing self-described components via a design process which involves assembly and production steps, after which many of the finished products are exported. However, high reliability and stability must be guaranteed, because customers require high-precision components at the time of manufacturing. In the aircraft parts industry, the mass production of high-value-added parts is limited. Therefore, a small production scale depending on the part is used, as many types of conventional CNC lathe machines with X-axis and Z-axis as well as Z-axis and C-axis CNC milling are used. The parts also rely on high-pressure air to increase production. The most important factors are good stability during processing, as high-precision parts are required, as noted above. It was found that as the C-axis rotation speed increased, the diameter of the cutting tool decreased with a decrease in the surface roughness, while the workpiece rotation speed increased with an increase in the surface roughness.

A Study on the Machining Accuracy according to Vibration and Unbalance Decrease in Rotational Speed Domains of High Precision Machine Tools (정밀 공작기계의 회전 영역별 진동 및 불평형량 감소에 따른 가공 정밀도 영향에 관한 연구)

  • Son, Deok-Soo;Kim, Sang-Hwa;Park, Il-Hwan
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.12 no.2
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    • pp.121-126
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    • 2013
  • Precision machine tools for high dignity cutting are needed for efforts to improve machining accuracy. However, there are many factors to improve machining accuracy. This study investigated how machining accuracy changes when variation and unbalance amount in rotational speed domain is decreased. Machining accuracy of initial machine tools depends on manufacturing and assembly of parts such as bearing. And then, vibration and noise vary with volume of unbalance amount when it is rotation, so it effects unbalance amount. Also vibration and noise increased by unbalance shorten spindle's life and it especially makes worse boring accuracy. Therefore, this study studied the change of roundness and cylindricity of workpiece when it decreases variation and unbalance in rotational speed domain.

Inactivation of Mad2B Enhances Apoptosis in Human Cervical Cancer Cell Line upon Cisplatin-Induced DNA Damage

  • Ju Hwan Kim;Hak Rim Kim;Rajnikant Patel
    • Biomolecules & Therapeutics
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    • v.31 no.3
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    • pp.340-349
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    • 2023
  • Mad2B (Mad2L2), the human homolog of the yeast Rev7 protein, is a regulatory subunit of DNA polymerase ζ that shares sequence similarity with the mitotic checkpoint protein Mad2A. Previous studies on Mad2B have concluded that it is a mitotic checkpoint protein that functions by inhibiting the anaphase-promoting complex/cyclosome (APC/C). Here, we demonstrate that Mad2B is activated in response to cisplatin-induced DNA damage. Mad2B co-localizes at nuclear foci with DNA damage markers, such as proliferating cell nuclear antigen and gamma histone H2AX (γ-H2AX), following cisplatin-induced DNA damage. However, unlike Mad2A, the binding of Mad2B to Cdc20 does not inhibit the activity of APC/C in vitro. In contrast to Mad2A, Mad2B does not localize to kinetochores or binds to Cdc20 in spindle assembly checkpoint-activated cells. Loss of the Mad2B protein leads to damaged nuclei following cisplatin-induced DNA damage. Mad2B/Rev7 depletion causes the accumulation of damaged nuclei, thereby accelerating apoptosis in human cancer cells in response to cisplatin-induced DNA damage. Therefore, our results suggest that Mad2B may be a critical modulator of DNA damage response.

Ruvbl1 is Essential for Ciliary Beating during Xenopus laevis Embryogenesis

  • Chan Young Kim;Hyun-Kyung Lee;Hongchan Lee;Hyun-Shik Lee
    • Development and Reproduction
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    • v.27 no.3
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    • pp.159-165
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    • 2023
  • The Ruvb-like AAA ATPase1 (Ruvbl1; also known as Pontin) is an evolutionary conserved protein belonging to the adenosine triphosphates associated with diverse cellular activities (AAA+) superfamily of ATPases. Ruvbl1 is a component of various protein supercomplexes and is involved in a variety of cellular activities, including chromatin remodeling, DNA damage repair, and mitotic spindle assembly however, the developmental significance of this protein is unknown and needs detailed investigation. We investigated the developmental significance of Ruvbl1 in multiciliated cells of the Xenopus laevis epidermis since ruvbl1 is expressed in the multiciliated cells and pronephros during X. laevis embryogenesis. The knockdown of ruvbl1 significantly impaired cilia-driven fluid flow and basal body polarity in the X. laevis epidermis compared to control embryos, but did not affect cilia morphology. Our results suggest that Ruvbl1 plays a significant role in embryonic development by regulating ciliary beating; however, further investigation is needed to determine the mechanisms involved.