• Genomics & Informatics
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• 제7권4호
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• pp.203-207
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• 2009

The target of rapamycin (TOR) signaling pathway conserved from yeast to human plays critical roles in regulation of eukaryotic cell growth. It has been shown that TOR pathway is involved in several cellular processes, including ribosome biogenesis, nutrient response, autophagy and aging. However, due to the functional diversity of TOR pathway, we do not know yet some key effectors of the pathway. To find unknown effectors of TOR signaling pathway, we took advantage of a green fluorescent protein (GFP)-tagged collection of budding yeast Saccharomyces cerevisiae. We analyzed protein abundance changes by measuring the GFP fluorescence intensity of 4156 GFP-tagged yeast strains under inhibition of TOR pathway. Our proteomic analysis argues that 83 proteins are decreased whereas 32 proteins are increased by treatment of rapamycin, a specific inhibitor of TOR complex 1 (TORC1). We found that, among the 115 proteins that show significant changes in protein abundance under rapamycin treatment, 37 proteins also show expression changes in the mRNA levels by more than 2-fold under the same condition. We suggest that the 115 proteins indentified in this study may be directly or indirectly involved in TOR signaling and can serve as candidates for further investigation of the effectors of TOR pathway.

• Journal of Species Research
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• 제13권1호
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• pp.100-104
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• 2024

The purpose of this study was to isolate and identify wild yeasts from soil samples collected in Daegu and Cheongju city, Republic of Korea. To identify the wild yeast strains, pairwise sequence comparisons of D1/D2 region of the 26S rRNA gene sequence were done using Basic Local Alignment Search Tool (BLAST). The cell morphologies were observed by phase contrast microscope and assimilation test are done using API 20C AUX kit. All strains were assigned to the phylum Basidiomycota. Among 13 strains, 11 strains were previously reported, but two strains were unreported from the Republic of Korea. The two unrecorded yeast strains, GW1-3 and PG1-1-10C, belong to the genus Solicoccozyma (family Piskurozymaceae, order Filobasidiales, class Tremellomycetes). The two strains had oval-shaped and polar budding cells. This research showed the morphological and biochemical properties of the two unreported yeast species that had not officially reported in Korea.

• 한국임상수의학회지
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• 제31권2호
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• pp.156-158
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• 2014

오타리아 물개의 탈모성 피부염에서 Malassezia spp. 를 분리하였다. Sabouraud dextrose agar에서 성장한 집락형태는 유백색에서 황색조로 변연부는 매끄러웠다. 현미경 검경시 형태는 원형에서 실린더형이었고 증식을 지시하는 budding 형이 관찰되었다. 개의 외이염에서 분리한 Malassezia pachydermatis와 비교하기 위해 26s rRNA 염기서열 분석을 실시하여 99.9% 의 일치도를 보였다. Itraconazole pulse therapy는 매우 효과적이었고 재발하지 않았다. 본 증례는 국내 물개에서 발생한 Malassezia 피부염 최초 보고로 판단된다.

• Journal of Microbiology and Biotechnology
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• 제15권5호
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• pp.1146-1151
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• 2005

The yeast Candida albicans has a distinguishing feature, dimorphism, which is the ability to switch between two morphological forms: a budding yeast form and a multicellular invasive filamentous form. This ability has been postulated to contribute to the virulence of this organism. Previously, we reported that the yeast-to-hypha transition in this organism is suppressed by farnesoic acid, a morphogenic autoregulatory substance that accumulates in the medium as the cells proliferate. In this study, using a differential display reverse transcription polymerase chain reaction (DDRT-PCR) technique, we have identified several genes induced in C. albicans by farnesoic acid treatment. These observations indicate that farnesoic acid can alter the expressivity of multiple genes, including the DNA replication machinery and cell-cycle-control proteins.

• Molecules and Cells
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• 제26권3호
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• pp.299-307
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• 2008

To characterize gene expression that is dependent on the strength of calorie restriction (CR), we obtained transcriptome at different levels of glucose, which is a major energy and carbon source for budding yeast. To faithfully mimic mammalian CR in yeast culture, we reconstituted and grew seeding yeast cells in fresh 2% YPD media before inoculating into 2%, 1%, 0.5% and 0.25% YPD media to reflect different CR strengths. We collected and characterized 160 genes that responded to CR strength based on the rigorous statistical analyses of multiple test corrected ANOVA (adjusted p value < 0.1 or raw p value < 0.0031) and Pearson correlation (|r| > 0.7). Based on the individual gene studies and the GO Term Finder analysis of 160 genes, we found that CR dose-dependently and gradually increased mitochondrial function at the transcriptional level. Therefore, we suggest these 160 genes are markers that respond to CR strength and that might be useful in elucidating CR mechanisms, especially how stronger CR extends life span more.

• Journal of Species Research
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• 제13권2호
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• pp.142-146
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• 2024

The purpose of this study was to isolate and identify wild yeasts from soil collected in Daegu City and Cheongyang County, Republic of Korea. Among 11 strains isolated in this study, nine strains were previously reported and two strains were unreported in Republic of Korea. To identify wild yeast strains, pairwise sequence comparisons of the D1/D2 region of the 26S rRNA gene sequence were done using Basic Local Alignment Search Tool (BLAST). The cell morphologies were observed by phase contrast microscope and assimilation test are done using API 20C AUX kit. All strains were assigned to the phylum Basidiomycota. Of the two unrecorded yeast strains, CY-9-10C belongs to the genus Mrakia (family Mrakiaceae, order Cystofilobasidiales, class Tremellomycetes) and PG3-4-10C belongs to the genus Slooffia (family Chrysozymaceae, order Microbotryomycetes incertae sedis, class Microbotryomycetes). Both strains had oval-shaped and polar budding cells. This research described the morphological and biochemical properties of the two unreported yeast species that had not officially reported in Korea.

• Journal of Microbiology
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• 제39권4호
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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.

• Journal of Microbiology and Biotechnology
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• 제29권1호
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• pp.141-150
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• 2019

The spatial landmark protein Bud8 plays a crucial role in bipolar budding in the budding yeast Saccharomyces cerevisiae. The unconventional yeast Yarrowia lipolytica can also bud in a bipolar pattern, but is evolutionarily distant from S. cerevisiae. It encodes the protein YALI0F12738p, which shares the highest amino acid sequence homology with S. cerevisiae Bud8, sharing a conserved transmembrane domain at the C-terminus. Therefore, we named it YlBud8. Deletion of YlBud8 in Y. lipolytica causes cellular separation defects, resulting in budded cells remaining linked with one another as cell chains or multiple buds from a single cell, which suggests that YlBud8 may play an important role in cell separation, which is distinct from the function of Bud8 in S. cerevisiae. We also show that the YlBud8-GFP fusion protein is located at the cell membrane and enriched in the bud cortex, which would be consistent with a role in the regulation of cell separation. The coiled-coil domain at the N-terminus of YlBud8 is important to the correct localization and function of YlBud8, as truncated proteins that do not contain the coiled-coil domain cannot rescue the defects observed in $Ylbud8{\Delta}$. This finding suggests that a new signaling pathway controlled by YlBud8 via regulation of cell separation may exist in Y. lipolytica.

• Molecules and Cells
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• 제41권1호
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• pp.35-44
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• 2018

Mitochondria are responsible for supplying of most of the cell's energy via oxidative phosphorylation. However, mitochondria also can be deleterious for a cell because they are the primary source of reactive oxygen species, which are generated as a byproduct of respiration. Accumulation of mitochondrial and cellular oxidative damage leads to diverse pathologies. Thus, it is important to maintain a population of healthy and functional mitochondria for normal cellular metabolism. Eukaryotes have developed defense mechanisms to cope with aberrant mitochondria. Mitochondria autophagy (known as mitophagy) is thought to be one such process that selectively sequesters dysfunctional or excess mitochondria within double-membrane autophagosomes and carries them into lysosomes/vacuoles for degradation. The power of genetics and conservation of fundamental cellular processes among eukaryotes make yeast an excellent model for understanding the general mechanisms, regulation, and function of mitophagy. In budding yeast, a mitochondrial surface protein, Atg32, serves as a mitochondrial receptor for selective autophagy that interacts with Atg11, an adaptor protein for selective types of autophagy, and Atg8, a ubiquitin-like protein localized to the isolation membrane. Atg32 is regulated transcriptionally and post-translationally to control mitophagy. Moreover, because Atg32 is a mitophagy-specific protein, analysis of its deficient mutant enables investigation of the physiological roles of mitophagy. Here, we review recent progress in the understanding of the molecular mechanisms and functional importance of mitophagy in yeast at multiple levels.

• Journal of Species Research
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• 제9권3호
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• pp.198-203
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• 2020

The goal of this study was to isolate and identify wild yeasts from soil samples. The 15 wild yeast strains were isolated from the soil samples collected in Pocheon city, Gyeonggi Province, Korea. Among them, four yeast stains were unrecorded, and 11 yeast stains were previously recorded in Korea. To identify wild yeasts, microbiological characteristics were observed by API 20C AUX kit. Pairwise sequence comparisons of the D1/D2 domain of the 26S rRNA were performed using Basic Local Alignment Search Tool(BLAST). Cell morphology of yeast strains was examined by phase contrast microscope. All strains were oval-shaped and polar budding and positive for assimilation of glucose, 2-keto-ᴅ-gluconate, N-acetyl-ᴅ-glucosamine, ᴅ-maltose and ᴅ-saccharose (sucrose). There is no official report that describes these four yeast species: one strain of the genus Kodamaea in the family Metschnikowiaceae and three strains of the Hannaella in the family Bulleribasidiaceae. Kodamaea ohmeri YI7, Hannaella kunmingensis YP355, Hannaella luteola YP230 and Hannaella oryzae YP366 were recorded in Korea, for the first time.