• 미생물학회지
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• 제40권2호
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• pp.178-182
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• 2004

Saccharomyces cerevisiae의 class I 키틴생합성 효소인 Chsl의 활성측정법을 적용하여 Aspergillus nidulans의 영양균사에서 class I 키틴생합성 효소인 ChsC의 활성측정을 시도하였다. 그 결과, A. nidulans의 class I 키틴생합성효소도 효모류의 경우와 마찬가지로 트립신 처리에 의하여 활성화되는 효소전구체형태(zymogenic form)로 존재함을 알 수 있었다. 그리고 A. nidulans 야생형의 class I 키틴 생합성 효소활성은 트립신 처리에 의하여 6배 가량 증가되었다. 반면, chsC 유전자가 파괴된 돌연변이주는 트립신 처리에 의하여 효소활성이 증가되지 아니하였을 뿐만 아니라, 효소활성의 수준도 트립신을 처리하지 아니한 야생형의 class I 키틴생합성 효소활성과 거의 동일한 수준이었다. 따라서, 트립신을 처리하여 측정한A. nidulans 야생형의 class I 키틴생합성 효소활성 값에서 트립신을 처리하지 아니한 야생형의 class I 키틴생합성 효소활성을 제외한 값이 A. nidulans 야생형의 ChsC 효소활성임을 알았다. 이러한 조건을 토대로 영양균사 생장과정 동안 ChsC의 효소활성을 측정한 결과, chsC 유전자의 발현양상과 유사하게 액체배양상태의 영양균사가 무성분화능을 획득하는 시기로 알려진 시간대에 효소의 활성이 증가하였다. 이러한 결과는 이미 보고된 바와 같이 chs 유전자가 A. nidulans의 영양균사 생장에 관여함을 시사하고 있다.

• 한국토양비료학회지
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• 제46권3호
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• pp.182-186
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• 2013

In this study, we examined the chemical properties and microbial community characteristics in 25 controlled horticultural soils (CHS) sampled from Gyeongnam Province by fatty acid methyl ester (FAME) method. The electrical conductivity of watermelon CHS was significantly (p < 0.05) higher than those of red pepper CHS, pumpkin CHS, and strawberry CHS. The amounts of total FAMEs, total bacteria, gram-negative bacteria, gram-positive bacteria, and fungi were significantly (p < 0.05) higher in red pepper CHS than those in strawberry CHS and pumpkin CHS. In addition, higher (p < 0.05) ratios of cy19:0 to $18:1{\omega}7c$ were detected in tomato CHS than those in watermelon CHS, pumpkin CHS, and red pepper CHS. This implied that microbial communities of tomato CHS were stressed more than other species of cultivation soils. Actinomycetes community in red pepper CHS was significantly (p < 0.05) higher than those in tomato CHS, strawberry CHS, and watermelon CHS. Differences in soil microbial community composition were highly associated with cultivated crop species which might result from the management inputs such as fertilizer, herbicide, and irrigation.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1995년도 한국생물과학협회 학술발표대회
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• pp.83-83
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• 1995

The three chitin synthetases of Saccharomyces cerevisiae, Chs1, Chs2, and Chs3, participate in septum and cell wall formation of vegetative cells and in wall morphogenesis of conjugating cells and spores. Because of the differences in the nature and in the time of execution of their functions, the synthetases must be specifically and individually regulated. The nature of that regulation has been investigated by measuring changes in the levels of the three synthetases and of the messages of the three corresponding gnes, CDSI, CHS2, and CAL1/CSD2/DITl0l(referred to below as CAL1), during the budding cycles. For Chs1 and Chs3, posttranslational regulation, probably by activation of latent forms, appears to be predominant. Since Chs2, like Chs1, is found in the cell in the zymogenic form, a posttranslational activation step appears to be necessary for this synthetase also. The regulation mechanism was investigated to search the relationship of CAL1, CAL2 and CALJ which is involved in Chs3 activity us ing different assay methods other than previous one. Treatment of Chs3-containing membranes with detergents drastically reduced the enzymatic activity. Activity could, however, be restored by subsequent incubation with trypsin or other pro teases in the presence of UDPGlcNAc. Experiments wi th mutants in the three genes invoIved in Chs3 activity-CAL1, CAL2, and CALJ-showed that only CAL1 and CALJ are required for the proteaseelicited (zymogenic) activity. It is concluded that Chs3 IS a zymogen and that the CAL2 product funct ions as its activator.ivator.

• 환경생물
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• 제23권4호
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• pp.335-342
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• 2005

We cloned seven genes encoding chitin synthases (CHSs) by PCR amplification from genomic DNAs of four strains of the genus Sporobolomyces and of Bensingtonia subrosea using degenerated primers based on conserved regions of the CHS genes. Though amino acid sequences of these genes were shown similar as 176 to 189 amino acids except SgCHS2, DNA sequences were different in size, which was due to various introns present in seven fragments. Alignment and phylogenetic analysis of their deduced amino acid sequences together with the reported CHS genes of basidiomycetes separated the sequences into classes I, II and III. This analysis also permitted the classification of isolated CHSs; SgCHS1 belongs to class I, BsCHS1, SaCHS1, SgCHS2, SpgCHS1, and SsCHS1 belong to class II, and BsCHS2 belongs to class III. The deduced amino acid sequences involving in class II that were discovered from five strains were also compared with those of other basidiomycetes by CLUSTAL X program. The bootstrap analysis and phylogenetic tree by neighbor-joining method revealed the taxonomic and evolutionary position for four strains of the genus Sporobolomyces and for Bensingtonia subrosea which agreed with the previous classification. The results clearly showed that CHS fragments could be used as a valuable key for the molecular taxonomic and phylogenetic studies of basidiomycetes.

• Journal of Microbiology and Biotechnology
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• 제12권6호
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• pp.943-949
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• 2002

It has been proposed that chitin synthase 3 (CHS3)-nediated chitin synthesis during the vegetative cell cycle is regulated by chitin synthase 4 (CHS4) of Saccharomyces cerevisiae. To investigate direct protein-protein interaction between the coding products of these two genes, a domain of Chs3p that is responsible for interaction with Chs4p was identified, using the yeast two-hybrid system. This domain of 54 amino acids, termed MIRC3-4 (Maximum Interacting Region of Chs3p with Chs4p), is well conserved among CHS3 homologs of various fungi. Some mutations in MIRC3-4 resulted in a decrease in the enzymatic activity and chitin contents. Chs3p carrying those mutations exhibited weak interactions with Chs4p, when assayed by the yeast two-hybrid system. Surprisingly, all the mutants were sensitive to Calcofluor regardless of changes in enzymatic activities or chitin contents. This report deals with a core region in MIRC3-4 that affects the interaction with Chs4p.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2000년도 추계학술발표대회
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• pp.39-45
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• 2000

It hab been proposed that CHS3-mediated chitin synthesis during the vegitative cell cycle is regulated by CHS4. To investigate direct protein-protein interaction between their coding products, we used yeast two hybrid system and found that a domain of Chs3p was responsible for interaction with Chs4p. This domain, termed MIRC3-4 (maximum interacting region of chs3p with chs4p), spans from 647 to 700 residues. It is well conserved among CHS3 homologs of various fungi such as Candida albicans, Emericella nidulans, Neurospora crassa, Magnaporthe grisea, Ustilago maydis, Glomus versiforme, Exophiala dermatitidis, Rhizopus microsporus. A series of mutaion in the MIRC3-4 resulted in no appearance of chitin ring at the early G 1 phase but did not affect chitin synthesis in the cell wall after cytokinesis. Absence of chitin ring could be caused either by delocalization of Chs3p to the septum or by improper interaction with Chs4p. To discriminate those two, not mutually exclusive, alternatives, mutants cells were immunostained with Chs3p-specific antibody. Some exhibited localization of chs3p to the septum, while others failed. These results indicate that simultaneous localization and activation Chs3p by Chs4p is required for chitin ring synthesis.

• Journal of Microbiology and Biotechnology
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• 제13권2호
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• pp.263-268
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• 2003

Recently, we identified a domain, termed MIRC3-4, for the protein-protein interaction between yeast chitin synthase 3 (CHS3) and chitin synthase 4 (CHS4). In this study, the functional roles of MIRC3-4 were examined at the G1 phase and cytokinesis of the cell cycle by Calcofluor staining and FISH. Some mutations in MIRC3-4 resulted in disappearance of the chitin ring in the early G1 phase, but did not affect chitin synthesis in the cell wall at cytokinesis. The chitin distribution in chs4 mutant cells indicated that CHS4 was involved in the synthesis of chitinring in the G1 phase and in the synthesis of cell wall chitin after cytokinesis, suggesting that Chs4p regulates chitin synthase 3 activity differently in G1 and cytokinesis. Absence of the chitin ring could be caused either by delocalization of Chs3p to the bud-neck or by improper interaction with Chs4p. When mutant cells were immunostained with a Chs3p-specific antibody to discriminate between these two alternatives, the mutated Ch3p was found to localize to the neck in all MIRC3-4 mutants. These results strongly irdicate that Chs4p regulates Chs3p as an activator but not a recruiter.

• 한국균학회지
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• 제26권3호통권86호
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• pp.354-360
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• 1998

여름 느타리 Pleurotus sajor-caju로부터 Chitin synthase(CHS) gene 특이 primer를 이용한 PCR을 통해 3개의 DNA 단편을 분리하여 cloning하였다. 분리된 DNA 단편들을 기존에 보고된 CHS 유전자들과의 염기서열을 분석한 결과, 이들 DNA 단편들 3개가 모두 CHS 유전자의 단편임을 확인하였고, 또한 이들은 각각 서로 다른 종류의 CHS 유전자들임을 알 수 있었다. 한편, RT-PCR 방법을 이용하여 분리된 유전자의 발현 실험을 실시해본 결과, 이들중 하나인 PsCHS3 유전자는 갓과 균사에서만 발현되는 기관특이 발현 특성을 보였으며, 또한 이 유전자는 상처 처리에 의해 그 발현이 증가되는 것을 확인하였다. 이러한 실험결과로 볼 때 p. sajor-caju의 경우, 다른 균류들의 경우처럼 다양한 기능을 가진 여러 종류(최소 3종류)의 CHS 유전자를 보유하고 있으며, 이들 각각은 다른 기관, 또는 다른 생육 단계에 작용하고 있을 것으로 생각되고, 특히 병 방어 기작에도 관여할 것으로 추측되어진다.

• Journal of Photoscience
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• 제7권2호
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• pp.53-58
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• 2000

We performed the cloning of a chalcone synthase (CHS) gene, the key enzyme in the anthocyanin biosynthesis, from the cDNA library constructed with grape suspension cells irradiated UV-B. The PCR fragment was used to cloning the CHS gene. One CHS cDNA clone containing an open reading frame and a partial stilbene synthase (STS)cDNA, the stilbene-type phytoalexin, were isolated. The CHS cDNA clone (VCHS) showed 87% sequence homology with VvCHS (V.vinifea) and 72.3% identity with VSTSY(V.vinifea). its amino acid sequences were longer than any other CHS genes as 454 residues. Two genes were weakly expressed in white light irradiated cells, but highly induced in UV-B irradiated condition during 32 hours. Interestingly, the STS was quickly and abundantly expressed from 2 hours when supplemented with jasmonic acid (JA) and the maximum expression was observed at 4 hours and then gradually decreased. But, the additional UV-B or white light quickly degraded the STS expression than only JA treated grape suspension cells. The CHS also was rapidly induced with JA and the synergistical effect was observed at the addigional light treatment of UV-B or white light. These results are indicated that CHS and STS have different response mechanisms against the environmental stresses.

• 동의생리병리학회지
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• 제20권3호
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• pp.710-718
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• 2006

The present study was carried out to investigate the effect of Chihyo-san (CHS) administration on asthma induced by Alum/OVA treatment in the mice. In CHS-treated animal group, lung weight, which was increased after asthma induction, was significantly decreased, and total number of cells in the lung, peripheral lymph node (PLN) and spleen tissue was significantly decreased in CHS-treated group compared to the asthma control group. The number of immune cells including natural killer (NK) cells in asthmatic animals was largely regulated by CHS treatment, showing a similar pattern as that of CsA-treated positive control group. Levels of mRNAs encoding inflammatory cytokines IL-5, IL-13, $TNF-{\alpha}$, and eotaxin were determined by RT-PCR in the lung tissue and showed decreases in CHS-treated group to the similar levels of CsA-treated control group, Histamine level in the serum was significantly lower in CHS-treated group than asthma-induced control group. Both haematoxylin and eosin staining and Masson's trichrome staining results showed decreased number of inflammatory cells, reduced immune cell infiltration, and normalized epithelial cell layering in the bronchial tissue of CHS-treated mouse group. Thus, the present findings suggest that CHS may be useful for protecting bronchial tissues from consistent inflammatory damages that occur in asthma patients.