• Title/Summary/Keyword: Budding Yeast

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Heterogeneous interaction network of yeast prions and remodeling factors detected in live cells

  • Pack, Chan-Gi;Inoue, Yuji;Higurashi, Takashi;Kawai-Noma, Shigeko;Hayashi, Daigo;Craig, Elizabeth;Taguchi, Hideki
    • BMB Reports
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    • v.50 no.9
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    • pp.478-483
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    • 2017
  • Budding yeast has dozens of prions, which are mutually dependent on each other for the de novo prion formation. In addition to the interactions among prions, transmissions of prions are strictly dependent on two chaperone systems: the Hsp104 and the Hsp70/Hsp40 (J-protein) systems, both of which cooperatively remodel the prion aggregates to ensure the multiplication of prion entities. Since it has been postulated that prions and the remodeling factors constitute complex networks in cells, a quantitative approach to describe the interactions in live cells would be required. Here, the researchers applied dual-color fluorescence cross-correlation spectroscopy to investigate the molecular network of interaction in single live cells. The findings demonstrate that yeast prions and remodeling factors constitute a network through heterogeneous protein-protein interactions.

Otitis Externa caused by Malassezia furfur in a Miniature Pig (애완 미니 돼지의 Malassezia furfur에 의한 외이도염 증례)

  • Han, Jae-Ik;Na, Ki-Jeong
    • Journal of Veterinary Clinics
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    • v.26 no.3
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    • pp.303-305
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    • 2009
  • A 7-month-old, female miniature pig was presented with excessive cerumen and pruritus. Greasy brown cerumen in both exteranal ear canal and sporadic head shaking were observed in the physical examination. Numerous budding yeasts in the cerumen were examined on microscopic examination. For species identification, PCR-RFLP using incubated colony on modified Dixon's medium was performed and finally, causative yeast was identified as M. furfur.

Msi1-Like (MSIL) Proteins in Fungi

  • Yang, Dong-Hoon;Maeng, Shinae;Bahn, Yong-Sun
    • Mycobiology
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    • v.41 no.1
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    • pp.1-12
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    • 2013
  • Msi1-like (MSIL) proteins, which are eukaryote-specific and contain a series of WD40 repeats, have pleiotropic roles in chromatin assembly, DNA damage repair, and regulation of nutrient/stress-sensing signaling pathways. In the fungal kingdom, the functions of MSIL proteins have been studied most intensively in the budding yeast model Saccharomyces cerevisiae, an ascomycete. Yet their functions are largely unknown in other fungi. Recently, an MSIL protein, Msl1, was discovered and functionally characterized in the pathogenic yeast Cryptococcus neoformans, a basidiomycete. Interestingly, MSIL proteins appear to have redundant and unique roles in both fungi, suggesting that MSIL proteins may have evolutionarily divergent roles in different parts of the fungal kingdom. In this review, we will describe the current findings regarding the role of MSIL proteins in fungi and discuss future directions for research on this topic.

Role of CaBud6p in the Polarized Growth of Candida albicans

  • Song Yun-Kyoung;Kim Jeong-Yoon
    • Journal of Microbiology
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    • v.44 no.3
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    • pp.311-319
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    • 2006
  • Bud6p is a component of a polarisome that controls cell polarity in Saccharomyces cerevisiae. In this study, we investigated the role of the Candide albicans Bud6 protein (CaBud6p) in cell polarity and hyphal development. CaBud6p, which consists of 703 amino acids, had 37% amino-acid sequence identity with the Bud6 protein of S. cerevisiae. The homozygous knock-out of CaBUD6 resulted in several abnormal phenotypes, such as a round and enlarged cells, widened bud necks, and a random budding pattern. In hypha-inducing media, the mutant cells had markedly swollen tips and a reduced ability to switch from yeast to hypha. In addition, a yeast two-Hybrid analysis showed a physical interaction between CaBud6p and CaAct1p, which suggests that CaBud6p may be involved in actin cable organization, like Bud6p in S. cerevisiae. Taken together, these results indicate that CaBud6 plays an important role in the polarized growth of C. albicans.

Effect of Environmental Stress on Morphological Change of an Extremely Cadmium-Tolerant Yeast, Hansenula anomala B-7

  • Huh, Nam-Eung;Choi, Nack-Shick
    • Journal of Microbiology and Biotechnology
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    • v.9 no.1
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    • pp.70-77
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    • 1999
  • An extremely cadmium-tolerant budding yeast, Hansenula anomala B-7 underwent a morphological switch in response to either heat shock treatment or cadmium stress, respectively. It exhibited a morphological transition from a unicellular yeast form to a pseudohyphae-like coagulation when subjected to prolonged heat shock treatment. In contrast, the yeast cells showed an irregularity in surface morphology when given thermal stress for a short time. Patterns of proteins expressed in the pseudohyphae-like cells demonstrated that several proteins were overexpressed while others were underexpressed in comparison with those prepared from the cells in the yeast form. It was a striking feature, however, that nearly 40% of the proteins extracted from the cells in the pseudohyphae form appeared to be composed of a single polypeptide. This polypeptide was apparently overexpressed during the pseudohyphae phase and its molecular weight was estimated to be 58 kDa according to SDS-PAGE analysis. However, a significant level of the protein was not observed in the cells before transition to pseudohyphae. The architecture of the cell shape was also damaged when incubated in a medium containing more than 1,000 ppm (8.9mM) of cadmium ions, although able to proliferate at a slow rate. However, the irregularity in the cell morphology exerted either by the brief heat shock treatment or by the cadmium stress with the high concentrations of the metal ions was not repaired, even though the damaged cells were allowed to grow for sufficient time in fresh, cadmium-free medium.

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A New Function of Skp1 in the Mitotic Exit of Budding Yeast Saccharomyces cerevisiae

  • Kim, Na-Mil;Yoon, Ha-Young;Lee, Eun-Hwa;Song, Ki-Won
    • Journal of Microbiology
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    • v.44 no.6
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    • pp.641-648
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    • 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.

Mycological Characteristics of Nine Unrecorded Yeasts from Flowers in the Orchard of Yesan-gun, Chungcheongnam-do and Hanbat Arboretum in Daejeon City, Korea

  • Han, Sang-Min;Hyun, Se-Hee;Shin, Ja-Won;Kim, Ha-Kun;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.42 no.3
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    • pp.231-234
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    • 2014
  • Six unrecorded yeasts, Cryptococcus festucosus 41-3, Cryptococcus heveanensis 56-4, Debaryomyces nepalensis 95-4, Issatchenkia occidentalis 142-1, Dioszegia zsoltii 39-1, and Kwoniella europala 47-2 were screened from 108 yeasts isolated from flowers and fruits in orchards of Yesan-gun, Chungcheongnam-do, Korea. The morphological and cultural characteristics of these unrecorded yeasts were investigated. They had various shapes, including ellipsoidal, globose, and oval, and also had budding mode in vegetable reproduction, except I. occidentalis 142-1 (fission mode). K. europaea 47-2 only formed pseudomycelium. D. zsoliti 39-1 did not grow in yeast extract-malt extract medium, potato dextrose medium, and vitamin-free medium. C. festucosus 41-3 grew well in 5% NaCl-containing yeast extract-peptone-dextrose medium and had a growth pH range of 7.0~10.0. Three unrecorded yeasts Ogataea polymorpha HB45-1, Rhodotonula hinnulla HB62-2, and Cryptococcus rajasthanensis HB80-4 were screened from 51 yeasts isolated from flowers in Hanbat arboretum in Daejeon city, Korea. They were globose in shape and did not form pseudomycelium. In addition, O. polymorpha HB45-1 and C. rajasthanensis HB80-4 had budding mode in vegetable reproduction. All of them grew well in vitamin-free medium and C. rajasthanesis HB80-4 also grew in 50% glucose and 5% NaCl-containing YPD medium.

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.

Hop2 and Sae3 Are Required for Dmc1-Mediated Double-Strand Break Repair via Homolog Bias during Meiosis

  • Cho, Hong-Rae;Kong, Yoon-Ju;Hong, Soo-Gil;Kim, Keun Pil
    • Molecules and Cells
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    • v.39 no.7
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    • pp.550-556
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    • 2016
  • During meiosis, exchange of DNA segments occurs between paired homologous chromosomes in order to produce recombinant chromosomes, helping to increase genetic diversity within a species. This genetic exchange process is tightly controlled by the eukaryotic RecA homologs Rad51 and Dmc1, which are involved in strand exchange of meiotic recombination, with Rad51 participating specifically in mitotic recombination. Meiotic recombination requires an interaction between homologous chromosomes to repair programmed double-strand breaks (DSBs). In this study, we investigated the budding yeast meiosis-specific proteins Hop2 and Sae3, which function in the Dmc1-dependent pathway. This pathway mediates the homology searching and strand invasion processes. Mek1 kinase participates in switching meiotic recombination from sister bias to homolog bias after DSB formation. In the absence of Hop2 and Sae3, DSBs were produced normally, but showed defects in the DSB-to-single-end invasion transition mediated by Dmc1 and auxiliary factors, and mutant strains failed to complete proper chromosome segregation. However, in the absence of Mek1 kinase activity, Rad51-dependent recombination progressed via sister bias in the $hop2{\Delta}$ or $sae3{\Delta}$ mutants, even in the presence of Dmc1. Thus, Hop2 and Sae3 actively modulate Dmc1-dependent recombination, effectively progressing homolog bias, a process requiring Mek1 kinase activation.

Isolation and characterization of four unrecorded wild yeasts from the soils of Republic of Korea in winter

  • Yuna Park;Soohyun Maeng;Sathiyaraj Srinivasan
    • Journal of Species Research
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    • v.12 no.3
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    • pp.197-202
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    • 2023
  • The purpose of this study was to isolate and identify wild yeasts from the soil collected in Gwangju and Pocheon City, Gyeonggi Province, Republic of Korea. Among 10 strains, six strains were already reported, but four strains were unrecorded in Republic of Korea. To identify wild yeast strains, pairwise sequence comparisons of the D1/D2 region of the 26S rRNA gene sequence were performed using Basic Local Alignment Search Tool (BLAST). The cell morphologies were observed by phase contrast microscope and assimilation tests were carried out using API 20C AUX kit. The 10 strains were assigned to the phyla Basidiomycota (8 strains) and Ascomycota (2strains). The unrecorded four yeast strains, NH33, NH19, NH20, and YP416, belong to the phylum Basidiomycota and the genera Buckleyzyma, Leucosporidium, Holtermanniales, and Mrakia, respectively. All strains had oval-shaped and polar budding cells. In this research, the morphological and biochemical properties of four unreported yeast species were characterized intensively, which were not officially reported in Korea.