• Molecules and Cells
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• 제38권3호
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• pp.243-250
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• 2015

Patterning of the polar axis during the early leaf developmental stage is established by cell-to-cell communication between the shoot apical meristem (SAM) and the leaf primordia. In a previous study, we showed that the DRL1 gene, which encodes a homolog of the Elongator-associated protein KTI12 of yeast, acts as a positive regulator of adaxial leaf patterning and shoot meristem activity. To determine the evolutionally conserved functions of DRL1, we performed a comparison of the deduced amino acid sequence of DRL1 and its yeast homolog, KTI12, and found that while overall homology was low, well-conserved domains were presented. DRL1 contained two conserved plant-specific domains. Expression of the DRL1 gene in a yeast KTI12-deficient yeast mutant suppressed the growth retardation phenotype, but did not rescue the caffeine sensitivity, indicating that the role of Arabidopsis Elongator-associated protein is partially conserved with yeast KTI12, but may have changed between yeast and plants in response to caffeine during the course of evolution. In addition, elevated expression of DRL1 gene triggered zymocin sensitivity, while overexpression of KTI12 maintained zymocin resistance, indicating that the function of Arabidopsis DRL1 may not overlap with yeast KTI12 with regards to toxin sensitivity. In this study, expression analysis showed that class-I KNOX genes were downregulated in the shoot apex, and that YAB and KAN were upregulated in leaves of the Arabidopsis drl1- 101 mutant. Our results provide insight into the communication network between the SAM and leaf primordia required for the establishment of leaf polarity by mediating histone acetylation or through other mechanisms.

• 한국컴퓨터정보학회논문지
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• 제14권3호
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• pp.193-207
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• 2009

세포 내에서 일어나는 단백질 신호 전달 과정은 단백질간의 상호작용을 통해 수행되고 조절된다. Yeast 상호작용 정보와 녹색형광단백질(GFP)을 이용하여 밝혀진 약 5,000여 개의 Yeast 단백질 위치정보를 이용하여 가중치를 부여하고 신호 전달경로 추출 및 예측을 위한 고성능 LocSPF 알고리즘을 최초로 제안하였다. 가중치 알고리즘에 의해 산출된 결과 중 의미 상관도가 높은 것을 채택한 후 KEGG에서 제공하는 신호전달 경로와 같은 신호전달 경로를 추출하는지 유사도 비교를 하였다. 한편 더 나아가 아직 실험을 통해 밝혀지지 않은 단백질 신호전달 경로를 예측하여 결과를 제시함으로써 본 연구를 통해서 알려지지 않은 새로운 신호전달 경로를 발견하거나 이전 경로에 참여하지 않은 단백질들을 발견할 수 있는 가능성을 제시 하였다.

• 한국식품과학회지
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• 제34권5호
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• pp.867-872
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• 2002

정미성이 높은 효모 추출물을 얻고자 각종 효소의 사용에 대한 최적 조합 및 공정법을 알아보기 위하여 맥주 폐효모박을 각종 효소로 처리하여 효모추출물 중의 정미성분(IMP, GMP 및 유리아미노산)을 측정하여 비교, 분석하였다. Glucanase(0.5%) 처리에 의한 효모추출물중의 조단백질 함량은 33.6% 이었다. Tunicase(1%) 28.0% 와 무처리구 21.1%에 비해 최고 1.6배의 증가를 보였다. 단백질 분해효소처리에 의한 조단백질의 함량은 bromelin(1%), protamex(1%) 처리에서 각각 30.8%, 29.8%로 무처리구에 비해 최고 1.4배의 증가를 보였다. 효소 복합처리에 의한 상승효과는 glucanase(0.5%)+protamex(1%) 처리구에서 조단백질의 함량이 34.4%로 나타나 glucanase 단독처리구의 33.6%보다 높은 함량을 나타냈다. IMP+GMP 총함량은 glucanase + phophodiesterase + adenyldeaminase (G+P+A) 혼합 처리구에서 1,066 mg/100 g, glucanase + ptotamex + phophodiesterase + adenyldeaminase (G+Pro+P+A) 혼합 처리구에서는 1,047 mg/100 g으로 비슷하였다. 유리아미노산의 함량은 protamex가 첨가된 G+Pro+P+A 혼합처리구에서 2,302 mg/100 g으로 가장 높게 나타났다. 따라서 IMP, GMP 및 유리아미노산의 함량을 모두 고려해 볼 때 세포벽 분해효소 (gulcanase 0.5%, 12시간), 단백질 분해효소 (protamex 1%, 3시간), 핵산 분해효소 (phosphodiesterase 0.1%, 3시간) 및 핵산 전이효소 (adenyldeaminase 1%, 1.5시간)를 순차적으로 적용시켜 가수분해시키는 것이 효모 추출물의 정미성을 높이는 최적의 효소 복합 사용공정으로 판단되었다.

• Journal of Microbiology
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• 제38권2호
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• pp.113-116
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• 2000

Escherichia coli F plasmid partition apparatus is composed of two trans-acting proteins (SopA and SopB) and one cis-acting DNA sequence (sopC). The SopB-sopC complex has been suggested to serve a centromere-like function through its interaction with chromosomally encoded proteins which remain to be identified. In this paper, we are introducing a new yeast 1.5-hybrid system which assembles the two-hybrid and one-hybrid system as a mean to find and additional component of the F plasmid partition system, interacting with DNA (sopC)-bound SopB protein. The results indicates that this system is a promising one, capable of selecting an interacting component.

• 미생물학회지
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• 제9권3호
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• pp.95-102
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• 1971

The growth characteristics of Candida tropicalis KIST 351 on gas oil substrate under different culture conditions were investigated and the preliminary animal feeding experiments using this yeast as a partial substitute of fish meal was also conducted. The yeast assimilates effectively n-paraffins in gas oil ranging from $C_{16}$ to $C_{16}$ with its maximum cell growth at $33^{\circ}C$ and pH 5.5 with aeration of 3 vvn and agitation of 900 rpm. The optimal concentrations of nitrogen sources, $HK_2PO_4$ and $Na_2HPO$ were 4, 2 and 0.5g/1, respectively. Ferrous sulfate, manganese sulfate and zinc sulfate showed positive effect to cell growth with the optimal range of 5-10 ppm. In the feeding experiment with 3 and 5% incorporation of the gas oil grown yeast, neither adverse effects on growth of chicks nor toxic effect were observed. Protein content of the dried cell was 58.8% and its amino acid composition compared well with other single-cell protein products and FAO reference protein.

• 한국식품영양과학회지
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• 제26권6호
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• pp.1246-1251
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• 1997

When the membrane protein fraction of mating type a cells of heterobasidiomycetous yeast R. toruloides was phosphorylated in vitro, two phosphorylated proteins of 72Kd and 57Kd were detected on SDS-polyacryamide gel. The phosphorylation reaction was inhibited by rhodotorucine A(Rh. A) which is a sexual pheromone secreted by mating type A cells. The inhibition of phosphorylation by Rh. A was dependent on $Ca^{2+}$, and independent on $Mg^{2+}$ or calmodulin. When adding trigger peptidase(TPase) inhibitor, antipain, no inhibition of phosphory was observed. Also, by adding the trysin-digested product of Rh. A, the phosphorylation was inhibited as the action of Rh. A. From these results, it is expected that the inhibition of membrane protein phosphorylation should be caused by the digested product of Rh. A with TPase.

• KSBB Journal
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• 제13권3호
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• pp.325-330
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• 1998

The methylotrophic yeast, Pichia pastoris, is known to be a potential host to offer many advantages for production of recombinant proteins. Fermentation parameters were optimized to enhance the heterologous ${\beta}$-galactosidase productivity with P. pastoris. Optimum concentration of methanol, used as inducer, was observed to be 8 g/L and the extent of repression of AOX1 promoter by glycerol was lower than by glucose. The degradation of the gene product ${\beta}$-galactosidase by protease was inhibited as the pH increased from 5 to 8 and the yeast extract(1%) as nitrogen source increased expression level 4 times higher compared to yeast nitrogen base(1%) as nitrogen source increased expression level 4 times higher compared to yeast nitrogen base(1%). Induction method, in which methanol is just added to fermentation medium without centrifugation, was found to be as much effective as the one with centrifugation.

• BMB Reports
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• 제34권2호
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• pp.139-143
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• 2001

Thioredoxin (Trx) is a redox protein possessing conserved sequence Cys-Gly-Pro-Cys in ail organisms. Trx acts as an electron donor of many proteins including thioredoxin peroxidase (TPx). Yeast Trx 2 has two redox active cysteine residues at positions 31 and 34. To investigate the redox activity of each cysteine, we generated mutants C31S, C34S, and C31S/C34S using site directed mutagenesis and examined the redox activity of Trx variants as an electron donor for yeast TPx enzymes. None of the three Cysmutated Trx proteins was active as a redox protein in the 5', 5'-dithiobis-(2-dinitrobenzoic acid) reduction under the condition of the presence of NADPH and thioredoxin reductase, and in the thioredoxin dependent peroxidase activity of yeast TPx II. C34S enhanced the glutamine synthetase protection activity of yeast TPx I, even though 100 times more protein was needed to exhibit the same activity to WT. The formation of a mixed disulfide intermediate between Trx and TPx II subunits was analyzed by SDS-PAGE. The mixed dieter form of TPx II was found only for C34S. These results suggest that Cys-31 more effectively acts as an electron donor for TPx enzymes.

• Animal cells and systems
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• 제12권3호
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• pp.143-148
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• 2008

Heterogeneous nuclear ribonucleoprotein E1(hnRNP E1) is one of the primary pre-mRNA binding proteins in human cells. It consists of 356 amino acid residues and harbors three hnRNP K homology(KH) domains that mediate RNA-binding. The hnRNP E1 protein was shown to play important roles in mRNA stabilization and translational control. In order to enhance our understanding of the cellular functions of hnRNP E1, we searched for interacting proteins through a yeast two-hybrid screening while using HeLa cDNA library as target. One of the cDNA clones was found to be human ribosomal protein L18a cDNA(GenBank accession number BC071920). We demonstrated in this study that human ribosomal protein L18a, a constituent of ribosomal protein large subunit, interacts specifically with hnRNP E1 in the yeast two-hybrid system. Such an interaction was observed for the first time in this study, and was also verified by biochemical assay.

• BMB Reports
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• 제50권9호
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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.