• Molecules and Cells
• /
• v.21 no.2
• /
• pp.251-260
• /
• 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.

• Journal of Microbiology
• /
• v.44 no.6
• /
• pp.641-648
• /
• 2006

We previously reported that Skp1, a component of the Skp1-Cullin-F-box protein (SCF) complex essential for the timely degradation of cell cycle proteins by ubiquitination, physically interacts with Bfa1, which is a key negative regulator of the mitotic exit network (MEN) in response to diverse checkpoint-activating stresses in budding yeast. In this study, we initially investigated whether the interaction of Skp1 and Bfa1 is involved in the regulation of the Bfa1 protein level during the cell cycle, especially by mediating its degradation. However, the profile of the Bfa1 protein did not change during the cell cycle in skp1-11, which is a SKP1 mutant allele in which the function of Skp1 as a part of SCF is completely impaired, thus indicating that Skp1 does not affect the degradation of Bfa1. On the other hand, we found that the skp1-12 mutant allele, previously reported to block G2-M transition, showed defects in mitotic exit and cytokinesis. The skp1-12 mutant allele also revealed a specific genetic interaction with ${\Delta}bfa1$. Bfa1 interacted with Skp1 via its 184 C-terminal residues (Bfa1-D8) that are responsible for its function in mitotic exit. In addition, the interaction between Bfa1 and the Skp1-12 mutant protein was stronger than that of Bfa1 and the wild type Skp1. We suggest a novel function of Skp1 in mitotic exit and cytokinesis, independent of its function as a part of the SCF complex. The interaction of Skp1 and Bfa1 may contribute to the function of Skp1 in the mitotic exit.

• Journal of Life Science
• /
• v.12 no.4
• /
• pp.452-459
• /
• 2002

Neutrophil apoptosis is a constitutive process that can be enhanced or delayed by various stimuli. In this study, effect of brefeldin A (BFA), which affects biological process of secretion, on constitutive and delayed apoptosis of neutrophils was investigated. Neutrophil apoptosis was determined after culturing for 20 hr in vitro by morphological changes, annexin V staining and DNA electrophoresis. BFA increased the constitutive apoptotic rate of neutrophils in dose-dependent manner. The delay of apoptosis induced by granulocyte macrophage-colony stimulating factor and lipopolysaccharide was also blocked by 10 $\mu$M of BFA. However, this effect of BFA was less marked when neutrophils were treated with dexamethasone, interleukin-8, or dibutyryl-cAMP. Moreover, the delay of neutrophil apoptosis induced by rottlerin, a specific inhibitor of protein kinase C-$\delta$ was significantly abrogated by BFA. Although BFA-induced apoptosis was not blocked by the caspase-3 inhibitor, zDEVD-fmk, expression levels of myeloid cell leukemia-1 (Mcl-1) were down-regulated by BFA. These results suggest that derangement of vesicular protein transport may be involved in the apoptosis of neutrophils, and that the action of BFA on apoptosis is dependent on changes in the expression of Mcl-1.

• Journal of Microbiology
• /
• v.39 no.4
• /
• pp.286-292
• /
• 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.

• BMB Reports
• /
• v.35 no.5
• /
• pp.518-523
• /
• 2002

Nordihydroguaiaretic acid (NDGA), an inhibitor of lipoxygenase, inhibits the secretion of proteins and causes the redistribution of resident Golgi proteins into the endoplasmic reticulum (ER). In this study, the effect of NDGA on lipoprotein lipase (LPL) secretion was investigated in 3T3-L1 adipocytes, and compared with those of brefeldin A (BFA), a well-known fungal metabolite that exhibits similar ER-Golgi redistribution. Both BFA and NDGA blocked secretions of LPL. In the presence of BFA, the active and dimeric LPL was accumulated in adipocytes. After endoglycosidase H (endo H) digestion, the proportion of LPL subunits with partially endo H-sensitive oligosaccharide was significantly increased with BFA. However, in the presence of NDGA, the cellular LPL became inactive, and only the endo H-sensitive fraction of the LPL subunit was observed. An increase of the aggregated forms was observed in the fractions of the sucrose-density gradient ultracentrifugation. These properties of LPL in the NDGA-treated cells were similar to those of LPL that is retained in ER, and the effects of NDGA could not be reversed by BFA. These results indicate that the inhibitory mechanism of NDGA on the LPL secretion is functionally different from the ER-Golgi redistribution that is induced by BFA.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.51 no.3
• /
• pp.259-268
• /
• 2006

Miniature inverted transposable elements (MITEs) are abundant genomic components in plant including rice. MITE-transposon display (MITE-TD) is an Amplified Fragment Length Polymorphism (AFLP)-related technique based on MITE sequence. In this study, we used the MITE-AFLP for the analysis of diversity and relation-ship of the 114 japonica accessions. Of the several MITEs, the mPing family was applied to detect polymorphisms based on PCR amplification. The BfaI adaptor primer and the specific primer derived from mPing terminal inverted repeat (TIR) region were used to PCR amplification of 114 accessions. Nine primer pairs produced a total of 160 polymorphic bands. PIC values of the polymorphic bands generated by nine primer pairs ranged from 0.269 (BfaI + ACT) to 0.426 (BfaI + T). Each accession revealed a distinct fingerprint with two primer combinations, BfaI + G and BfaI + C. Cluster analysis using marker-based genetic similarity classified 114 accessions into five groups. MITE-AFLP markers were genetically mapped using a population of 80 BILs (BC1F7) derived from a cross between the rice accessions, Milyang 23 and Hapcheonaengmi 3. Eight of the markers produced with the primer pair BfaI + 0 were mapped on chromosomes 1, 2, 4, 5, 7, and 9. Considering that one MITE-AFLP marker on chromosome 7 was tightly linked to the Rc gene, the MITE-AFLP markers will be useful for gene tagging and molecular cloning.

• Proceedings of the Microbiological Society of Korea Conference
• /
• 2006.05a
• /
• pp.135-135
• /
• 2006

• Environmental Engineering Research
• /
• v.25 no.1
• /
• pp.104-113
• /
• 2020

Adsorptive breakthrough modelling is essential for design of a sorption packed bed. In this work, breakthrough modelling of the furfural uptake in bagasse fly ash (BFA) packed bed has been performed. Effect of various parameters like bed height (Z = 15-60 cm), flow rate (Q = 0.02-0.04 L/min) and initial furfural concentration (C_o = 50-200 mg/L) on the breakthrough curve of furfural sorption in a BFA packed bed have been studied. Enhanced breakthrough performance was observed for the higher value of Z, and lower values of C_o and Q. For C_o = 100 mg/L, packed bed operated at Q = 0.03 L/min and Z = 60 cm was found to have lowest adsorbent utilization rate of 5.61 g/L with highest breakthrough volume of 14.67 L. Bed depth service time and Thomas models well represented the experimental data points under all experimental conditions. It can be concluded that BFA can be utilized efficiently in continuous system for the removal of furfural. Overall, more than 99% of furfural was adsorbed in BFA packed bed at experimental conditions.

• Analytical Science and Technology
• /
• v.7 no.4
• /
• pp.435-440
• /
• 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.

• Journal of Periodontal and Implant Science
• /
• v.34 no.1
• /
• pp.177-193
• /
• 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.