• Title/Summary/Keyword: yeast form

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SCG10, a Microtubule-Destabilizing Factor, Interacts Directly with Kinesin Superfamily KIF1A Protein in Brain (Kinesin superfamily KIF1A와 결합하는 미세소관 불안정화 단백질 SCG10의 규명)

  • Moon, Il-Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.19 no.7
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    • pp.859-865
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    • 2009
  • Microtubules, a major cytoskeleton, form parallel arrays in the axon and are oriented with their plus ends toward the cell periphery. Kinesin superfamily proteins (KIFs) are the molecular motors acting in the microtubule-based motilities of organelles in cells. Here, we used the yeast two-hybrid system to identify the protein that interacts with the coiled-coil domain of KIF1A and found a specific interaction with microtubule-destabilizing factor SCG10. SCG10 bound to the amino acid residues between 400 and 820 of KIF1A, but not to other KIFs in the yeast two-hybrid assay. The coiled-coil domain of SCG10 is essential for interaction with KIF1A. In addition, this specific interaction was also observed in the Glutathione S-transferase pull-down assay. An antibody to SCG10 specifically co-immunoprecipitated KIF1A associated with SCG10 from mouse brain extracts. These results suggest that KIF1A motor protein transports SCG10-containing vesicles along microtubules in neurons.

Na/K-ATPase beta1-subunit associates with neuronal growth regulator 1 (NEGR1) to participate in intercellular interactions

  • Cheon, Yeongmi;Yoo, Ara;Seo, Hyunseok;Yun, Seo-Young;Lee, Hyeonhee;Lim, Heeji;Kim, Youngho;Che, Lihua;Lee, Soojin
    • BMB Reports
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    • v.54 no.3
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    • pp.164-169
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    • 2021
  • Neuronal growth regulator 1 (NEGR1) is a GPI-anchored membrane protein that is involved in neural cell adhesion and communication. Multiple genome wide association studies have found that NEGR1 is a generic risk factor for multiple human diseases, including obesity, autism, and depression. Recently, we reported that Negr1-/- mice showed a highly increased fat mass and affective behavior. In the present study, we identified Na/K-ATPase, beta1-subunit (ATP1B1) as an NEGR1 binding partner by yeast two-hybrid screening. NEGR1 and ATP1B1 were found to form a relatively stable complex in cells, at least partially co-localizing in membrane lipid rafts. We found that NEGR1 binds with ATP1B1 at its C-terminus, away from the binding site for the alpha subunit, and may contribute to intercellular interactions. Collectively, we report ATP1B1 as a novel NEGR1-interacting protein, which may help deciphering molecular networks underlying NEGR1-associated human diseases.

Screening of Anti-inflammatory Compound-producing Wild Yeasts and Their Microbiological Characteristics (항염증 물질 생산 능력이 우수한 야생효모의 선별 및 이들의 균학적 특성)

  • Bae, Sang-Min;Han, Sang-Min;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.45 no.3
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    • pp.212-223
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    • 2017
  • To screen for potent anti-inflammatory compound-producing yeasts, we evaluated nitric oxide production inhibitory activities in RAW 264.7 macrophage cells using cell-free extracts from 182 non-pathogenic yeasts. Rhodotorula graminis YJ36-1 and Meyerozyma guilliermondii YJ34-2 showed high inhibitory activities of 57.4% and 47.0%, respectively. The microbiological characteristics of these yeasts were investigated. Rhodotorula graminis YJ36-1 formed ascospores and pseudomycelium. This species grew well at $25^{\circ}C$ in yeast extract-peptone-dextrose (YPD) medium, vitamin-free medium, and 5% NaCl-containing YPD medium. Meyerozyma guilliermondii YJ34-2 was an asporogenous yeast and did not form pseudomycelium. This strain also grew well at $30^{\circ}C$ in YPD medium, vitamin-free medium, and 5% NaCl-containing YPD medium.

Effects of High Dietary Levels of Selenium-Enriched Yeast and Sodium Selenite on Macro and Micro Mineral Metabolism in Grower-Finisher Swine

  • Kim, Y.Y.;Mahan, D.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.2
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    • pp.243-249
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    • 2001
  • Thirty six barrows with an initial body weight of 28 kg were used to determine the effect of two dietary Se sources and a wide range of Se levels encompassing 0.3, 1.0, 3.0, 5.0, 7.0, and 10.0 mg/kg Se. The organic Se form was a Se-enriched yeast product, whereas the inorganic Se source was sodium selenite. The experiment was a $2{\times}6$ RCB design conducted in three replicates. Each barrow was placed in an individual metabolism crate and provided their dietary treatment and water on an ad libitum basis for a minimum 2 wk period, whereupon feed intake was adjusted to a constant intake within replicate at approximately 90% of intake for a 4 d adjustment period. Urine and feces were subsequently collected for a 7 d period and analyzed for Se and minerals. The results demonstrated that urinary Se was approximately 25% higher when pigs were fed sodium selenite (p<0.01), whereas fecal Se was lower by 25% (p<0.01). Se retention tended to be higher when organic Se was provided (p>0.15). Urinary Se increased as dietary Se level increased for both Se sources but increased more and at a high rate when sodium selenite was fed resulting in an interaction response (p<0.01). Fecal Se increased linearly as the dietary level of both Se sources increased, but the fecal Se from organic Se increased at a faster rate resulting in an interaction response (p<0.01). Se retention increased linearly (p<0.01) as dietary Se increased for both Se sources. The apparent digestibility of Se increased by Se level when pigs were fed sodium selenite, but not when the organic Se source was provided resulting in an interaction response (p<0.05). Retention of consumed Ca, Zn increased when pigs were fed organic Se (p<0.05) whereas P and Na retention were higher when the inorganic Se was provided. Mineral retention was not affected by dietary Se level except P. These results suggest that Se excretion by urine was the main route of excretion when pigs were fed sodium selenite but the fecal route when Se-enriched yeast was provided. The excretion of Fe, Zn, Mn, and Cu via urine and feces was not affected by high dietary Se level or dietary Se sources.

Characterization of Protoplast Fusant between Killer Yeast and Alcohol-Fermenting Yeast (Killer 효모와 알콜 발효효모간의 원형질체 융합주의 특성)

  • 정기택;방광웅;김재근;송형익;정용진
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.55-64
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    • 1990
  • Cell volume and DNA contents of the fusants were similar to those of parents. Genetic stability of the fusants was increased when they were cultured on minimal medium (MM) rather than on complete medium (CM), and the fusants were stabilized by subculturing 7 generations each 7 day on MM agar. The finally selected fusants after being cultured for 6 months on CM were stable without segregation. The fusants could also form nuclein and ascospores, and show red and pink colors by the test of TTC colorization. Assimilability and fermentability of carbon sources of the fusants were similarto those of parents. The tolerance of KCl, NaCl, sodium propionate and cycloheximide showed the traits of one strain of parents. When the fusants were cultured for 72 hr and 60 hr in the medium containing 20% glucose and sucrose, respectively, the yield of ethanol for FWKS 260 was reached to 9.6 v/v% and 9.8v/v%, respectively. The sensitive strain Kyokai 7 was found to be killed entirely after cultivation of 48 hr by the killer toxin from the fusants. The recipient S 29 and Kyokai 7 were found to have neither L nor M dsRNA plasmid. However, K 52 and fusants had both L and M dsRNA plasmid of 4.7 kb and 2.5 kb, respectively. The curants treated by heat and cycloheximide did not contain M dsRNA plasmid, but had large amounts of L dsRNA plasmid of those of killer yeasts.

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Function and Oligomerization Study of the Leucine Zipper-like Domain in P13 from Leucania separata Multiple Nuclear Polyhedrosis Virus

  • Du, Enqi;Yao, Lunguang;Xu, Hua;Lu, Songya;Qi, Yipeng
    • BMB Reports
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    • v.40 no.2
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    • pp.232-238
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    • 2007
  • The p13 gene is uniquely present in Group II nucleopolyhedroviruses (NPVs) and some granuloviruses, but not in Group I NPVs. p13 gene was first described by our laboratory in Leucania separatamultiple nuclear polyhedrosis virus (Ls-p13) in 1995. However, the functions of Ls-P13 and of its homologues are unknown. When Ls-p13 was inserted into Autographa californica nucleopolyhedrovirus, a Group I NPV, polyhedra yield was inhibited. However, this inhibition was prevented when the leucine zipper-like domain of Ls-p13 was mutated. To determine the cause of this marked difference between Ls-P13 and leucine zipper mutated Ls-P13 (Ls-P13mL), oligomerization and secondary structure analyses were performed. High performance liquid chromatography and yeast two-hybrid assays indicated that neither Ls-P13 nor Ls-P13mL could form oligomers. Informatics and circular dichroism spectropolarimetry results further indicated marked secondary structural differences between Ls-P13 and Ls-P13mL. The LZLD of Ls-P13 has two extended heptad repeat units which form a hydrophobic surface, but it is short of a third hydrophobic heptad repeat unit for oligomerization. However, the mutated LZLD of Ls-P13mL lacks the above hydrophobic surface, and its secondary structure is markedly different. This difference in its secondary structure may explain why Ls-P13mL is unable to inhibit polyhedra yield.

Diagnostic test for bovine mastitis by the determination of ATP based on firefly bioluminescence (Bioluminescence 반응에 의한 ATP 측정을 이용한 젖소 유방염 진단에 관한 연구)

  • Kim, Tae-jong;Kim, Jong-bae;Lee, Seoung-bae;Jeon, Young-soo
    • Korean Journal of Veterinary Research
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    • v.29 no.3
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    • pp.383-391
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    • 1989
  • This study was carried out to diagnostic test for bovine mastitis by the determination of adenosine triphosphate (ATP) based on firefly bioluminescence. The results obtained are follow; 1. The infection rate of bovine mastitis investigated with 521 cows in 47 dairy farms were found to be 3.6% of clinical form and 44.1% of subclinical form according to the degree of infection. 2. The light yield produced in firefly bioluminescence system was proportional to the concentration of ATP giving stright line within the range of 100PM~1uM. 3. When the number of somatic cell in milk was determined by the ATP assay and compared with three conventional methods such Fossomatic, California mastatic test (CMT), and rolling ball viscometer (RBV), it was shown that r=0.92 for Fossomatic, 0.63 for CMT and 0.7 for RBV. 4. The microorganisms causing mastitis were isolated Staphylococcus sp. (53.3%), Streptococcus sp. (17.9%), Micrococcus sp. (13.5%), Gram negative bacilli (6.3%), Gram positive bacilli (5.5%) and Yeast-like fungi (5.4%). 5. The endogeneous ATP levels of bacteria in a raw milk determined by the firefly bioluminescence system and compared with the results of the conventional methods. The correlation was 0.88 for raw milk.

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Disruption of the Dual Specificity Kinase Gene Causes the Reduction of Virulence in Candida albicans (이중특이성 인산화 효소의 결손이 Candida albicans 병원성에 미치는 효과)

  • Park, Yun-Hee;Park, Hee-Moon
    • The Korean Journal of Mycology
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    • v.39 no.1
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    • pp.85-87
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    • 2011
  • The opportunistic human pathogen Candida albicans has the ability to convert from yeast-form to pseudohyphal or true hyphal form. The morphological transition is considered as an important virulence factor, because the decrease or lack in dimorphism causes the reduction of virulence. Our previous study revealed that the disruption of dual specificity kinase gene caused the reduction of dimorphism in C. albicans. Therefore we tested the effect of dual specificity kinase in virulence using mouse model. The mean survival time for kinase-defective strains was about 15 days in comparison with those of wild-type, 3.9 days. Moreover the fungal burden on kidneys for kinase-defective strains was decreased by ten-fold than that for wild-type. These results suggest possible involvement of dual specificity kinase in a novel signal transduction pathway for morphological transition and virulence of C. albicans.

A Survey on Microbial Contamination of Currently-Sold Drugs (I) -Bacterial contamination of marketed liquid- (시장의약품(市販醫藥品)의 미생물(微生物) 오염도(汚染度) 조사(調査) (제1보)(第1報) -시판내복액제(市販內服液劑)의 세균오염도(細菌汚染度)-)

  • Park, Young-Ju;Kim, Young-Il
    • Journal of Pharmaceutical Investigation
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    • v.3 no.4
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    • pp.5-15
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    • 1973
  • An investigation was carried out on a basis of the bacteriological examination with a view to detecting the degree of bacterial contamination for the 77 samples collected from the locally-sold liquid specialties. It's test period was 50 days from July 10 to August 30, 1971. Specially, the survey has put emphasis on the population of general bacteria and the identification of coli-form group, staphylococcus species, streptococcus species, bacillus species, fungi, and yeast species from liquid samples. The results obtained are summarized as follows; (1) For the 77 samples tested, the contamination of general bacteria was found out as minimun 0, i,e., maximum, $12{\times}10^4$ and the total average $45{\times}10^2$ per milliliter. (2) Although streptococcus species could not be detected with the samples, the contamination of the coli-form and staphylococcus species means the strong suggestion of the possibility of pathogenic bacterial contamination. (3) Specially, the products which stay in the neutral pH range and use suspending agents need to care for the microbial contamination in the manufacturing crocess. (4) It is thought necessary to perform the microbiological quality control in the liquid preparations only at least. (5) As the microbial contamination degree in the liquid decreases according to the elapse of time, the microbiological quality control will have to be carried out immediately after the completion of the manufacturing process in order to know the accurate degree. (6) The author thinks that the main reason of the microbial contamination in the liquid is the contamination during the manufacturing process. (7) For the purpose of prevention of the microbial contamination in liquid, therefore, it is more important to make efforts for the rationalization of manufacturing process, the improvement of equipment and environment, the specific training of workers for hygienic knowledges, etc. rather than the use of preservatives for the preparations.

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Morphological and Physiological Characteristics of Amanita hemibapha subsp. hemibapha (Berk. & Broome) Sacc. (달걀버섯의 형태 및 생리적 특성)

  • Sou, Hong-Duck;Hur, Tae-Chul;Joo, Sung-Hyun
    • The Korean Journal of Mycology
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    • v.37 no.1
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    • pp.41-48
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    • 2009
  • This study was conducted to investigate optimum cultural conditions for Amanita hemibapha isolated in Korea and its hyphae characteristics. Micrographs shows the presence of clamp connection. A. hemibapha grows as mycelial form(M-phase) 2-4 ${\mu}m$ and yeast-like form(Y-phase) 7-8 ${\mu}m$. The fungal spores were broadly elliptical and papillate, 8-11 ${\times}$ 6-9 ${\mu}m$ in size. The nucleotide sequence analysis of the ITS of nuclear ribosomal DNA from sporocarps and in-vitro-grown mycelium supported the fungal species is Amanita hemibapha. A. hemibapha showed sequence similarity in the ITS rDNA with A. caesarea(97.5) and A. jacksonii(98.5%) which are morphologically similar species to A. hemibapha. The optimal pH and temperature for mycelial growth of A. hemibapha were pH 6.0 and $28^{\circ}C$, respectively. The fungal species showed best growth in SYP and GYS medium. A. hemibapha grew well with mannitol and glucose as carbon sources and peptone as a nitrogen source.