• Title/Summary/Keyword: Bfa1p

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The Study of Bfa1pE438K Suggests that Bfa1 Control the MitoticExit Network in Different Mechanisms Depending on DifferentCheckpoint-activating Signals

  • Kim, Junwon;Song, Kiwon
    • Molecules and Cells
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    • v.21 no.2
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    • pp.251-260
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    • 2006
  • During mitosis, genomic integrity is maintained by the proper coordination of anaphase entry and mitotic exit via mitotic checkpoints. In budding yeast, mitotic exit is controlled by a regulatory cascade called the mitotic exit network (MEN). The MEN is regulated by a small GTPase, Tem1p, which in turn is controlled by a two-component GAP, Bfa1p-Bub2p. Recent results suggested that phosphorylation of Bfa1p by the polorelated kinase Cdc5p is also required for triggering mitotic exit, since it decreases the GAP activity of Bfa1p-Bub2p. However, the dispensability of GEF Lte1p for mitotic exit has raised questions about regulation of the MEN by the GTPase activity of Tem1p. We isolated a Bfa1p mutant, $Bfa1p^{E438K}$, whose overexpression only partially induced anaphase arrest. The molecular and biochemical functions of $Bfa1p^{E438K}$ are similar to those of wild type Bfa1p, except for decreased GAP activity. Interestingly, in $BFA1^{E438K}$ cells, the MEN could be regulated with nearly wild type kinetics at physiological temperature, as well as in response to various checkpoint-activating signals, but the cells were more sensitive to spindle damage than wild type. These results suggest that the GAP activity of Bfa1p-Bub2p is responsible for the mitotic arrest caused by spindle damage and Bfa1p overproduction. In addition, the viability of cdc5-2 ${\Delta}bfa1 $ cells was not reduced by $BFA1^{E438K}$, suggesting that Cdc5p also regulates Bfa1p to activate mitotic exit by other mechanism(s), besides phosphorylation.

Characterization of a Putative F-box Motif in Ibd1p/Bfalp, a Spindle Checkpoint Regulator of Budding Yeast Saccharomyces cerevisiae

  • Lee, Kyum-Jung;Hyung-Seo;Kiwon Song
    • Journal of Microbiology
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    • v.39 no.4
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    • pp.286-292
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    • 2001
  • During mitosis. the proper segregation of duplicated chromosomes is corrdinated by a spindle check-point. The bifurcated spindle checkpoint blocks cell cycle progression at metaphase by monitoring unattached kinetochores and inhibits mitotic exit in response to the misorientation of the mitotic spin- dle Ibd1p/Bfa1p is a spindle checkpoint regulator of budding yeast in the Bub2p checkpoint pathway for mitotic exit and its disruption abolishes mitotic arrest when proper organization of the mitotic spin-dls inhibited. Ibd1p/Bfa1p localizes to the spindle pole body, a microtublue-organizing center in yeast, and its overexpression arrests the cell cycle in 80% of cells with an enlarged budy at mitosis and in 20 % of cells with multiple buds. In this study, we found that the C-terminus of Ibd1p/Bfa1p phys-ically interacts with Skp1p, a key component of SCF (Skp1/cullin/F-box) complex for ubiquition-medi-ated proteolysis of cel cycle regulatores as well as an evolutionally conserved kinetochore protein for cell cycle progression. A putative F-box motif was found in the C-terminus of Ibd1p/Bfa1p and its function was investigated by making mutants of conserved residues in the motif. These Ibd1p/Bfa1p mutants of a putative F-box interacted with SKp1p in vitro by two-hybrid assays as wild type Ibd1p/Bfa1p. Also these Ibd1p/Bfa1p utants displayed the overexpression phenotypes of wild type Ibd1p, when over-expressed under inducible promoters . These results suggest that a putative F-box motif of Ibd1p/Bfa1p is not essential for the interaction with SKp1p and its function in mitotic exit and cytokinesis.

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Analysis of the Anionic Surfactants by Capillary Electrophoresis (모세관 전기영동 장치를 이용한 음이온계 계면활성제의 분석)

  • Jeong, Hyuk;Kim, Seung Sun;Lee, Byung Min;Kang, Ho-Cheol;Lee, Won;Kim, Hai-Dong
    • Analytical Science and Technology
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    • v.7 no.4
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    • pp.435-440
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    • 1994
  • Qualitative and quantitative analysis for the anionic surfactants used in the metal washing fluid (brand names are BFA and BCA) was performed by the capillary electrophoresis. Acetonitrile and sodium benzoate were mixed with the buffer solution which controlled at pH 10. Under the 18kV applied voltage, the electropherograms have shown the theoretical plates more than $10^4$. Determined as the concentration at the S/N~3, the typical detection limit was ~5 ppm and the calibration curves have shown the correlation coefficients higher than ~0.99. Based on these results, it was concluded that each components were octanoate, decanoate, dodecanoate, tetradecanoate, hexadecanoate and the relative ratio was 1.0 : 1.0 : 6.5 : 2.1 : 0.8 for the BFA.

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Effects of electrical stimulation on healing of endo-osseous titanium implants in circumferential defect (전기자극이 성견 골결손부에 매식된 임플란트 주위조직의 치유에 미치는 영향)

  • Shim, Jae-Chang;Kim, Young-Jun;Chung, Hyun-Ju;Kim, Ok-Su
    • Journal of Periodontal and Implant Science
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    • v.34 no.1
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    • pp.177-193
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    • 2004
  • Several experimental studies showed that the application of small amounts of electric current to bone stimulated osteogenesis at the site of the cathode and suggested that electrical currents promote osseointegration around dental implants. The purpose of this study was to determine the effect of direct microcurrent to endosseous titanium implants placed in bone defects. The right and left 2nd, 3rd and 4th mandibular premolars in ten mongrel dogs (15Kg of weight) were extracted. One monthe later, Ti-machined screw type implants(3.8 mm diameter x 8.5 mm length, $AVANA^{(R)}$, Ostem) were placed in surgically created circumferential defect area(width 5mm, depth 4mm). The implants were divided into three groups according to the treatment modalities: Control group- implants without electrical stimulation; Experimental group I- implants with allogenic demineralized freeze dried bone grafting; and Experimental group II-implants allogenic demineralized freeze dried bone grafting and electric stimulation. The animals were sacrificed in the 4th and 8th week after implant placement and un-decalcified specimens were prepared for histological and histometrical evaluation of bone-implant contact ratio (BIC) and bone formation area ratio (BFA) in defect area. Some specimens at 8 weeks after implantation were used for removal torque testing. Histologically, there was connective tissue infiltration in the coronal part of defect area in control and the experimental group I, whereas direct bone contact was found in the experimental group II without connective tissue invasion. Average BIC ratios at 4 weeks of healing were 60.1% in the experimental group II, 47.4% in the experimental group I and 42.7% in the control. Average BIC ratios at 8 weeks after implantation were 67.6% in the experimental group II, 55.9% in the experimental group I and 54.6% in the control. The average BFA ratio was 84.0% in the experimental group II, 71.8% in the experimental group I and 58.8% in the control at 4 weeks, and the BFA ratios were 89.6% in the experimental group II, 81.4% in the experimental group I and 70.5% in the control at 8 weeks after implantation. The experimental group II showed also significantly greater BIC and BFA ratios compared to the control and the experimental group I (p<0.05). The removal torque values at 8 weeks after implantation were 56 Ncm in the experimental group II, 49 Ncm in the experimental group I and 43 Ncm in the control. There was a statistically significant difference among 3 groups (p<0.05). These results suggest that electrical stimulation improve and accelerate bone healing around endosseous titanium implants in bone defect.

Evidence of an Alternative Route of Cellobiase Secretion in the Presence of Brefeldin A in the Filamentous Fungus Termitomyces clypeatus

  • Banik, Samudra Prosad;Pal, Swagata;Chowdhury, Sudeshna;Ghorai, Shakuntala;Khowala, Suman
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.412-420
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    • 2011
  • Secretion of cellobiase occurred in a brefeldin A (BFA) uninhibited manner in the filamentous fungus Termitomyces clypeatus. Fluorescence confocal microscopy revealed that application of the drug at a concentration of 50 ${\mu}g$/ml caused arrest of Spitzenkorper assembly at the hyphal tip. This resulted in greater than 30% inhibition of total protein secretion in the culture medium. However, the cellobiase titer increased by 17%, and an additional 13% was localized in the vacuolar fraction en route secretion. The secretory vacuoles formed in the presence of the drug were also found to be bigger (68 nm) than those in the control cultures (40 nm). The enzyme secreted in the presence and absence of BFA revealed a single activity band in both cases in native PAGE and had similar molecular masses (approx. 120 kDa) in SDS-PAGE. The BFA enzyme retained 72% of native glycosylation. It also exhibited a higher stability and retained 98% activity at $50^{\circ}C$, 93.3% activity at pH 9, 63.64% activity in the presence of 1M guanidium hydrochloride, and 50% activity at a glucose concentration of 10 mg/ml in comparison to 68% activity, 75% activity, 36% activity, and 19% activity for the control enzyme, respectively. The observations collectively aimed at the operation of an alternative secretory pathway, distinct from the target of brefeldin A, which bypassed the Golgi apparatus, but still was able to deliver the cargo to the vacuoles for secretion. This can be utilized in selectively enhancing the yield and stability of glycosidases for a successful industrial recipe.

Bisphenol-A Removal in Conventional Water Treatment Systems (정수처리공정에서 bisphenol-A의 제거에 관한 연구)

  • 김혜리;이윤진;박선구;남상호
    • Journal of Environmental Health Sciences
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    • v.30 no.1
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    • pp.59-64
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    • 2004
  • This study was carried out to investigate influencing factors of bisphenol A(BPA) removal characteristic in conventional water treatment systems to be connected with coagulation, sedimentation, filtration and disinfection. The result are summarized as follows; In BPA removal, optimal doses of PAC, alum, ferric chloride were 7.5 mg Al/L, 10.0 mg AI/L, 15.0 mg Fek. PAC was most effective coagulant to remove BPA. In coagulation process, BPA removal efficiency were increased about 2% by adjusting pH of raw water as 6. At temperature rise 1$0^{\circ}C$, BPA removal efficiency were increased 0.94%. but BPA removal efficiency in sand filtration process were under 1 %ie, so that BPA was almost not removed. At free chlorine dose 1, 2 mg/L, the reaction rate constant k in the BPA removal have been calculated to be 0.397, 0.953 min$^{-1}$ . At free chlorine dose 1, 2 mg/1-, degradation reaction of BPA was completed during 10 min and BFA removal efficiencies were 97.66, 99.99% at this time.

A Survey on Elementary School Childrens′ Awareness of and Preference for Kimchi (초등학생의 김치에 대한 의식과 선호 실태에 관한 연구)

  • 한재숙;김혜영;김정숙;서봉순;한준표
    • Korean journal of food and cookery science
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    • v.13 no.3
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    • pp.259-265
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    • 1997
  • The main purpose of this research is to provide a basic knowledge of Kimchi and to improve elementary school childrens' Kimchi intake. We researchers have done statistical analyses of 808 questionnaires completed by elementary school children and their mothers on their awareness of and preference for Kimchi. The results were as follows: About 80.2% of the children regarded Kimchi as one of our nutritious and traditional foods, and 83.8% of the subjects had an affirmative opinion of Kimchi intake. The most well known Kimchi is as follows: Baechu Kimchi, Kkaktugi, Nabak Kimchl, Chonggak Kimchi, Saaennip Kimchi and Buchu Kimchi. The children preferred the peculiar and refreshing taste of Kimchi. Their preference of Kimchi is in the following order: Baechu Kimchi, Skaktugi, Chonggak Kimchi, Nabak Kimchi and Oi Sobagi. And the children also preferred the stems of the cabbage. The children preferred properly fermented and freshly prepared Kimchi. Their favorite ingredients were red pepper powder, Korean radishes, pickled anchovies, sesame leaves, garlic and scallions. The children wanted Kimchi that is less hot and more sweet. The ingredients children wanted to add to Kimchi are pears, cuttlefish, oranges, apples and cucumbers.

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