• 제목/요약/키워드: Mcm complex

검색결과 14건 처리시간 0.025초

Biochemical Properties of the Minichromosomal Maintenance Complex after the Phosphorylation by Cdc7 Kinase

  • Lee, Joon-Kyu
    • Animal cells and systems
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    • 제10권1호
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    • pp.1-6
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    • 2006
  • Previous studies showed that Cdc7 kinase of Schizosaccharomyces pombe phosphorylated the minichromosome maintenance (Mcm) complex efficiently in the presence of spMcm10 protein. The biochemical properties of the phosphorylated Mcm complexes were examined to understand the activation mechanism of the Mcm complex by Cdc7 kinase. The phosphorylation of Mcm complex in the presence of spMcm10 by Cdc7 kinase did not affect the stability of the Mcm complex containing all six subunits, and the changes in the sedimentation properties were not observed after the phosphorylation. The reconstitution of the Mcm complex using the purified proteins showed that the phosphorylation of Mcm2 proteins did not affect the interactions between Mcm proteins. The phosphorylation of the Mcm2-7 complex at the same condition also did not activate the other biochemical activities such as DNA helicase and single stranded (ss) DNA binding activities. On the other hand, spMcm10 protein that was used for the stimulation of Mcm phosphorylation showed single stranded DNA binding activity, and inhibited the DNA helicase activity of the Mcm4/6/7 complex. These inhibitory effects were reduced by the addition of Cdc7 kinase, suggesting that the phosphorylation by Cdc7 kinase decreased the interactions between spMcm10 and the Mcm complex. Taken together, these results suggested that the phosphorylation by Cdc7 kinase alone is not sufficient for the remodeling and the activation of the Mcm complex, and the additional factors or the phosphorylations might be required for the activation of the Mcm complex.

Identification and characterization of a rice MCM2 homologue required for DNA replycation

  • Cho, Jae-Han;Kim, Ho-Bang;Kim, Hyung-Sae;Choi, Sang-Bong
    • BMB Reports
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    • 제41권8호
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    • pp.581-586
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    • 2008
  • The pre-replication complex (pre-RC), including the core hexameric MCM2-7 complex, ensures that the eukaryotic genome is replicated only once per cell division cycle. In this study, we identified a rice $\underline{m}ini\underline{c}hromosome$ $\underline{m}aintenance$ (MCM) homologue (OsMCM2) that functionally complemented fission yeast MCM2 (CDC19) mutants. We found OsMCM2 transcript expression in roots, leaves, and seeds, although expression levels differed slightly among the organs. Likewise, the OsMCM2 protein was ubiquitously expressed, but it was downregulated when nutritients were limiting, indicating that MCM2 expression (and therefore cell cycle progression) requires adequate nutrition. Yeast two-hybrid and GST pull-down assays demonstrated that OsMCM2 interacted with the COP9 signalosome 5 (CSN5). Taken as a whole, our results indicated that OsMCM2 functions as a subunit of the rice MCM complex and interacts with CSN5 during developmental regulation.

Adsorbate Interactions of Cu(II) Ion-Exchanged into Mesoporous Aluminosilicate MCM-41 Analyzed by Electron Spin Resonance and Electron Spin Echo Modulation

  • Kim, Jeong-Yeon;Yu, Jong-Sung
    • 한국자기공명학회논문지
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    • 제3권2호
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    • pp.109-126
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    • 1999
  • The location of Cu(II) exchanged into measoporous aluminosilicate MCM-41(AlMCM-41) material and its interaction with various adsorbate molecules were investigated by electron spin resonance and electron spin echo modulation spectroscopies. Cu(II) is fully coordinated to adsorbates in a wide open mesopore of AlMCM-41 for the formation of favorable complexes. It was found that in the fresh hydrated material, Cu(II) is octahedrally coordinated to six water molecules as evidenced by an isotropic room temperature ESR signal. This species is located in a cylindrical MCM-41 channel and rotates rapidly at room temperature. Evacuation at room temperature removes some of these water molecules, leaving the Cu(II) coordinated to less water molecules and anchored to oxygens in an MCM-41 channel wall. Dehydration at 450$^{\circ}C$ produces one Cu(II) species located on the internal wall of a channel, which is easily accessible to adsorbates. Adsorption of adsorbate molecules such as water, methanol, ammonia, pyridine, aniline, acetonitrile, benzene, and ethylene on a dehydrated Cu-AlMCM-41 material causes changes in the ESR spectrum of Cu(II), indicating the complex formation with these adsorbates. Cu(II) forms a complex with six molecules of methanol as evidenced by an isotropic room temperature ESR signal and ESEM analysis like upon water adsorption. Cu(II) also forms a square planar complex containing four molecules of N-containing adsorbates such as ammonia, pyridine and aniline based on resolved nitrogen superhyperfine interaction and their ESR parameters. However, Cu(II) forms a complex with six-molecules of acetonitrile based on ESR parameters. Only one molecule of benzene or ethylene is coordinated to Cu(II).

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Cupric Ion Species in Cu(II)-Exchanged Mesoporous MCM-41 Gallosilicate Determined by Electron Spin Resonance Studies

  • Kim, Jeong-Yeon;Yu, Jong-Sung
    • 한국자기공명학회논문지
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    • 제1권2호
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    • pp.126-140
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    • 1997
  • Mesoporous MCM-41 gallosilicate material was synthesized through shifting through shifting gallosilicate polymer equilibrium towards a MCM-41 phase by addition of acid. The location of Cu(II) exchanged into MCM-41 and its interaction with various adsorbate molecules were investigated by electron spin responance and electron spin echo modulation spectroscopies. It was found that in the fresh hydrated material, Cu(II) is octahedrally coordinated to six water molecules. This species is located in a cylindrical channel and rotates rapidly at room temperature. Evacuation at room temperature removes three of these water molecules, leaving the Cu (II) coordinated to three water molecules and anchored to oxygens in the channel wall. Dehydration at 45$0^{\circ}C$ produces one Cu (II) species located in the inner surface of a channel as evidenced by broadening of its ESR lines by oxygen. Adsorption of polar molecules such as water, methanol and ammonia on dehydrated CuNa-MCM-41 gallosilicate material causes changes in the ESR spectrum of Cu (II), indicating the complex formation with these adsorbates. Cu (II) forms a complex with six molecules of methanol as evidenced by an isotropic room temperature ESR signal and ESEM data like upon water adsorption. Cu(II) also forms a complex containing four molecules of ammonia based on resolved nitrogen superhyperfine interaction.

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MCM promoter에서 E-box와 E2F 결합부위가 전사활성에 미치는 영향 (Effect of E-box and E2F Binding Site on Transcriptional Activity in MCM Promoter)

  • 권현주
    • 생명과학회지
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    • 제14권5호
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    • pp.732-740
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    • 2004
  • MCM 단백질의 암세포내에서 과발현 되는 원인을 규명하기 위하여 mcm7, 2 promoter영역의 전사인자 결합 부위의 역할을 조사하였다. 암세포에서 promoter 활성은 정상세포보다 8배정도 높은 활성을 나타내 특유 전사인자의 존재를 시사하였다. Promoter영 역 에는 E2F 결합부위 와 Myc/Max/Mad단백질 결합 부위로 알려진 I-box가 존재한다. 이들 각각의 변이체 promoter를 작성하여 암세포와 정상세포에서 luciferase reporter 활성을 측정하는 것으로 각 영역의 역할을 검토하였다. 그 결과 정상세포 내에서는 강한 repressor존재 또는 활성을 negative로 조절 할 수 있는 complex의 존재로 promoter활성이 조절되는 반면 암세포 내에서는 I-box에 결합하여 promoter를 활성화시키는 특유 단백질의 존재가 시사되었다.

고속/고집적 ATM Switching MCM 구현을 위한 설계 Library 구축 밀 시험성 확보 (Generation of Testability on High Density /Speed ATM MCM and Its Library Build-up using BCB Thin Film Substrate)

  • 김승곤;지성근;우준환;임성완
    • 마이크로전자및패키징학회지
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    • 제6권2호
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    • pp.37-43
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    • 1999
  • 대용량, 고속 정보처리가 요구되는 시스템의 모듈은 데이터 처리의 고속성 및 회로의 고집적이 가능한 MCM의 형태로 구현되어 ATM, GPS 및 PCS 등의 분야에 광범위하게 응용되고 있다. 3개의 칩으로 구성되고 2.48 Gbps의 데이터 처리용량을 가지는 ATM Switching 모듈을 기판 Size 48$\times$48mm2, Cu/PhotoBCB를 이용한 10 Multi-Layer 그리고 491 Pin PBGA 형태의 MCM을 개발하였다. MCM 개발을 위해 요구되는 기술로는 고속신호 특성구현을 위해 Interconnect Characterization을 통한 기판/ 패키지의 설계 파라미터 추출, 고밀도 MCM 에서의 방열처리 그리고 MCM 개발의 가장 난점중의 하나인 시험성 확보를 들 수 있다. ATM Switching MCM 개발을 위해 MCM-D 기판에서의 Interconnect Characterization을 통한 신호지연, 비아특성, 신호간섭(Cross-talk) 파라미터 등을 추출하였다. 고집적 구조에서 15.6Watt의 방열처리를 위해 열 해석을 진행하고 기판에 열 비아 1.108개를 형성하고 패키지 전체에 $85^{\circ}C$ 이하 유지조건의 방열처리를 하였다. 마지막으로 시험성 확보를 위해 미세 간격 프로빙을 통한 기판 검증 및 복잡한 패키지/어셈블리 공정검증을 위해 Boundary Scan Test(BST)를 적용하여 효과적이고 비용 절감형의 제품을 개발하였다.

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Depletion of the Pre-RC Proteins Induces Chk1/Chk2 Independent Checkpoint Responses and Apoptotic Cell Death in HeLa Cells

  • Im, Jun-Sub;Lee, Joon-Kyu
    • Animal cells and systems
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    • 제11권2호
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    • pp.129-134
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    • 2007
  • The initiation of eukaryotic DNA replication requires assembly of the pre-replicative complex (Pre-RC) through the concerted action of Orc, Cdc6, Cdt1 and Mcm2-7 complex during G1 phase. The pre-RC assembly licenses individual replication origins for the initiation of DNA replication and sufficient number of the pre-RC is essential for proper progression of S phase. However, it is not well known how cells recognize the completion of the pre-RC assembly before G1-S transition. In order to understand the cellular responses to the defects in pre-RC assembly, we depleted the known components of pre-RC proteins using the small interference RNAs in HeLa cells. Although the defects of pre-RC assembly by the depletion of the pre-RC proteins such as Orc2, Cdt1, Mcm2 & Mcm10 did not elicit the activation of Chk1- or Chk2-dependent checkpoint pathways, these cells still showed significant decrease in the cellular level of Cdc25A proteins. These results suggests that a novel checkpoint pathway exist in HeLa cells, which is not dependent upon Chk1 or Chk2 proteins and play essential roles in the cellular responses to the defects in the pre-RC assembly. Also, among those four proteins tested in this study, the depletion of Mcm10 and Cdt1 proteins significantly increased the apoptotic cell death in HeLa cells, suggesting that these proteins not only play roles in the pre-RC assembly, but also are involved in the checkpoint responses to the defects in the pre-RC assembly.

백강균(Beauveria bassiana)의 균사체 최적 배양조건 및 효소활성 (Optimal Condition for Mycelial Growth of Beauveria bassiana and Its Extracellular Enzyme Activity)

  • 민응기;한영환
    • 미생물학회지
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    • 제38권1호
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    • pp.50-53
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    • 2002
  • 백강균(Beauveria bassiana DGUM 34001)은 $24^{\circ}C$의 온도와pH 7.0의 초기 pH에서 최적의 균사체 생육을나타내었다. 사용한 복합배지 중 mushroom complex배지(MCM)에서 가장 우수한 균사생육을 나타내었다. Czapek-Dox 한천배지를 최소배지로 각각 탄소원, 질소원 및 인산원의 영향을 조사한 결과, glucose, soytone 및 sodium phosphate ($NaH_2$$PO_4$)에서 가장 우수한 균사 생육을 보여주었다. MCM액체배지에서 균사 배양 후 세포외 효소 활성을 측정한 결과,$\alpha$-amylase, lipase, chitinase, CMCase및 pretense의 비효소활성은 각각 297.0, 0.058, 0.33, 0.21 및 22.8 units/mg protein이었다. Casein과 soluble chitosan을 첨가할 경우 세포외 분비 pretense와 chitinase의 효소 활성 이 증가되었다.

Evidence of DNA Replication Licensing and Paternal DNA Degradation by MCM7 and ORC2 in the Mouse One-cell Embryo

  • Kim, Chang Jin;Kim, Tae Hoon;Lee, Eun-Woo;Lee, Kyung-Bon
    • 대한의생명과학회지
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    • 제23권4호
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    • pp.372-379
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    • 2017
  • This study was investigated to test whether paternal DNA that was destined for degradation was properly licensed by testing for the presence of mini-chromosome maintenance protein (MCM) 7 and origin recognition complex (ORC) 2 in the paternal pronuclei. ORC2 is one of the first licensing protein to come on and MCM7 is one of the last licensing protein to come on. Zygotes were prepared by injection of control and treated sperm injection (ICSI). To control for DNA breakage, epididymal spermatozoa were treated with DNase I to fragment the DNA, then injected into oocytes. The presence of MCM7 and ORC2 in the pronuclei of mouse zygotes was tested by immunohistochemistry, just before the onset of DNA synthesis, at 5 h after fertilization, and after DNA synthesis began, at 9 h post fertilization. We found that in all cases, both MCM7 and ORC2 were present in both pronuclei at 5 h after sperm injection, just before DNA synthesis began. This indicates that no matter how extensive the DNA damage, recruitment of licensing proteins to the origins of replication was not inhibited. Sperm DNA fragmentation does not prevent licensing of DNA replication origins. Furthermore, the embryo recognizes DNA that is damaged by nucleases. Our data indicate that the one-cell embryo does harbor a mechanism to prevent the replication of severely damaged DNA from spermatozoa, even though the embryos do not undergo classical apoptosis.

Mega cisterna magna in bipolar mood disorder: a case report

  • Yazici, Esra;Kose, Sefanur;Gunduz, Yasemin;Kurt, Elif Merve;Yazici, Ahmet Bulent
    • Journal of Yeungnam Medical Science
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    • 제39권1호
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    • pp.58-61
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    • 2022
  • Mega cisterna magna (MCM), one of the members of the Dandy-Walker complex, is a developmental malformation of the posterior fossa that is larger than 10 mm but morphologically does not affect the vermis and cerebellar hemispheres. Reports of psychiatric disorders associated with this anomaly are rare. We present the case of a patient with MCM who presented with a psychotic manic attack and was diagnosed with bipolar disorder. A 28-year-old female, single housewife, university graduate, presented with irritability, decreased sleep and appetite, distraction, and agitation. The patient also had a delusion of reference. In the clinical follow-up, an increase in energy and an increase in the amount of speech were observed. Her neurological examination was normal, and cranial magnetic resonance imaging revealed an MCM. The relationship and clinical significance of MCM with psychosis and mood disorders have not yet been fully elucidated. It is not known whether this association is accidental or based on etiological commonality. The purpose of this case report is to review the relationship between the cerebellum and psychiatric symptoms and to contribute to the literature.