Gibberellic Acid 의 작용기작에 관한 연구 ( 3 ) . cyclic AMP 에 의한 단백질의 생합성 및 인산화 반응의 조절

심웅섭,노광수,문혜연 ( Woong Seop Sim,Kwang Soo Roh,Hye Yeon Moon ) 생화학분자생물학회 1983 BMB Reports Vol.16 No.1

In order to investigate cAMP`s involvement in the action mechanisms of gibberellic acid, the effects of cAMP on the biosynthesis and phosphorylation of proteins in the germinating maize seeds were studied. cAMP promoted the biosynthesis and phosphorylation of proteins. In the presence of actinomycin D, the stimulation of protein biosynthesis by cAMP was continuous during the germination period, while the protein phosphorylation was increased at the 3-day point after germination by CAMP. The stimulatory effect of cAMP on the protein biosynthesis was greater than that on the prntein phosphorylation during the early stages of germination. On the other hand, this pattern is reversed during the latter period. The above mentioned results are consistent with the facilitation effects of GA₃ on the protein biosynthesis and phosphorylation. Therefore, it seems to be that cAMP is related to the action mechanisms of GA₃.

Studies on the Mechanisms of Gibberellic Acid Action(III). Regulation of Protein Biosynthesis and Phosphorylation by cyclic AMP

심웅섭,노광수,문혜연,Sim, Woong-Seop,Roh, Kwang-Soo,Moon, Rye-Yeon 생화학분자생물학회 1983 한국생화학회지 Vol.16 No.1

cAMP가 $GA_3$의 작용기작에 관여하는 지를 밝히기 위해 단백질의 생합성과 인산화반응에 미치는 cAMP의 효과를 구명하였던바, 정상상태에서의 단백질의 생합성 및 인산화 반응은 cAMP에 의해 모두 촉진되었다. Transcription이 억제된 상태에서는 단백질의 생합성은 전 발아기간 중 대조구에 비해 촉진되었으나 인산화 반응은 발아 3일 후 cAMP에 의해 증가되었다. 또한 cAMP는 발아 초기에는 단백질의 인산화 반응 보다는 생합성을 촉진시키고, 발아가 진행됨에 따라 단백질의 생합성 보다는 인산화 반응을 촉진시킨다. 이러한 결과는 단백질의 생합성 및 인산화반응에 미치는 $GA_3$의 촉진효과와 동일한 점으로 보아 cAMP는 $GA_3$의 작용기작과 밀접한 관계가 있는 것으로 추정된다. In order to investigate cAMP's involvement in the action mechanisms of gibberellic acid, the effects of cAMP on the biosynthesis and phosphorylation of proteins in the germinating maize seeds were studied. cAMP promoted the biosynthesis and phosphorylation of proteins. In the presence of actinomycin D, the stimulation of protein biosynthesis by cAMP was continuous during the germination period, while the protein phosphorylation was increased at the 3-day point after germination by cAMP. The stimulatory effect of cAMP on the protein biosynthesis was greater than that on the prtein phosphorylation during the early stages of germination. On the other hand, this pattern is reversed during the latter period. The above mentioned results are consistent with the facilitation effects of $GA_3$ on the protein biosynthesis and phosphorylation. Therefore, it seems to be that cAMP is related to the action mechanisms of $GA_3$.

Effects of Cyclic AMP on the Activities of Ribosome and Elongation Factors EF-1 and EF-2

심웅섭,문혜연,Sim, Woong-Seop,Moon, Hye-Yeon 생화학분자생물학회 1986 한국생화학회지 Vol.19 No.3

단백질의 생합성에 필요한 여러가지 components의 활성에 미치는 cAMP 의 효과를 조사하였다. cAMP로 처리된 ribosome의 활성은 대조구에 비하여 27% 증가하였으며 EF-1 과 EF-2의 활성은 cAMP에 의하며 각각 17%와 20% 증가 하였다. The effects of cAMP on the activities of the various components necessary for protein biosynthesis were investigated. 1. The rate of protein biosynthesis was increased about 27% by ribosomes treated with cAMP. 2. The activities of elongation factors EF-1 and EF-2 were considerably stimulated by exogenous cAMP.

Gibberellic Acid 의 작용기작에 (作用機作) 관한 연구 - 1. CA3 에 의한 단백질의 생합성 및 인산화반응의 조절

심웅섭(Woong Seop Sim),노광수(Kwang Soo Roh) 한국식물학회 1979 Journal of Plant Biology Vol.22 No.4

As a part of the studies on the regulatory mechanism of gene expression by GA_3, the effects of GA_3 on the protein biosynthesis and phosphorylation in maize seedlings were investigated. 1. The optimum concentration of GA_3 for the stimulation of the protein biosynthesis was 0.3mM. 2. The protein biosynthesis was remarkably increase by GA_3 during the germination. The reason for the decrease in the protein biosynthesis by 48hrs. after germination seems to be a staggered gene expression, and/or increases in protease and RNase activities. 3. The ratio of the amount of the newly synthesized protein in germination seeds treated with GA_3 to the amount of proteins secreted into the endosperm was similar to that ratio in control. According to this result, it seems that GA_3 stimulates only the expression of certain definite genes. 4. By the treatment with GA_3, the rates of biosynthesis and phosphorylation of proteins were increased up to about 1.5 times during germination and 6 times by 72hrs. after germination, respectively. The ratio of the total soluble proteins to the phosphoproteins considerably increased in teh early germination stage (24hrs.) but decreased after 24hrs. According to the above mentioned results, the stimulation of the phosphorylation of proteins by GA_3 seems to be attributed to the increases in the activities of protein kinases.

Gibberellic Acid 의 작용 기작에 관한 연구 - 2. Actinomycin D 처리시 GA3 에 의한 단백질의 생합성 및 인산화반응의 (燐酸化反應) 조절

심웅섭(Woong Seop Sim),박홍덕(Hong Duok Park),노광수(Kwang Soo Roh),문혜연(Hye Yeon Moon) 한국식물학회 1982 Journal of Plant Biology Vol.25 No.1

As a part of the studies on the regulatory mechanism of gene expression by gibberellic acid, the effects fo GA_3 on the protein biosynthesis and phosphorylation in maize seedlings were investigated in the presence of actinomycin D. The activites of protein biosynthesis and phosphorylation in germination seeds treated with GA_3 were greater than these of the control at the 3-day point after germination. It is assumed that the enhancement of protein biosynthesis by GA_3 in the presence of actinomycin D is due to the effects of GA_3 on the translational processes in which protein is produced from the mRNA synthesized previously.

Transformation of Potato (Solanum tuberosum L.) Tuber Cells with Agrobacterium tumefaciens and Ti Plasmid DNA

정상호,심웅섭,Chung, Sang-Ho,Sim, Woong-Seop 생화학분자생물학회 1987 한국생화학회지 Vol.20 No.4

본 연구에서는 Agrobacterium tumefaciens 및 Ti-plasmid에 의한 감자 괴경세포의 형질전환 여부를 조사하여 다음과 같은 결과를 얻었다. Agrobacterium tumefaciens KU-12의 감염에 의하여 감자 괴경의 원반에 tumor가 형성되었으며, 이 원반을 호르몬이 없는 배지에서 배양한 결과 tumor에서 완전한 식물체가 재생되었다. 또한 tumor에서 재생된 식물체 내에는 lysopine dehydrogenase(LpDH) 활성이 있음을 확인하였다. 감자의 원형질체에 Ti-plasmid DNA(pTi-12)를 처리하여 호르몬이 결여된 배지에서 배양한 결과, 165개의 callus를 얻었으며, 그 중 일부에서는 뿌리가 형성되었다. 뿌리를 형성한 callus와 형성하지 않은 callus을 임의로 선택하여 LpDH 활성을 조사한 결과 둔 경우 모두 octopine을 합성함으로 형질전환체임을 확인하였다. 형질 전환된 식물의 genome내의 T-DNA 부위를 분석하기 위하여 DNA-DNA 혼성화실험을 성시한 결과, pTi-12 DNA내에 존재하는 T region내에는 Sma I fragment들 중 분자량이 134 kb, 37 kb 및 4.75 kb인 것이 완전히 내포되어 있으며, T region의 양쪽 끝은 분자량이 각각 9.7 kb와 4.55 kb인 Sma I fragment에 연관되어 있음을 알 수 있었고, 또한 T-DNA에 내포된 Sma I fragment들은 pTi-12 DNA 분자내에서 서로 연결된 상태로 존재하고 있음을 확인하였다. In the present study, transformation of potato tuber cells by Agrobacterium tumefaciens and Ti plasmid treatment was studied. The results are as follows: Tumors were induced in potato tuber discs by infection with Agrobacterium tumefaciens KU-12. Whole plants were regenerated from the tumors on hormone-free medium. It was identified that the lysopine dehydrogenase (LpDH) activity was present in the tumor-derived plants. A total of 165 calli were obtained from Ti-plasmid DNA(pTi-12)-treated potato protoplasts on hormone-free medium and a few of them formed a root. Each of the non-rooted and rooted callus was randomly selected and tested for the presence of LpDH activity. Both of them were ocotopine positive, indicating that they were transformants. In order to analyze the T-DNA region maintained in transformed plant genome, the DNA-DNA hybridization experiment was performed. As a result, it was found that Sma I fragments of Ti plasmid (pTi-12) 13.4 kb, 3.7 kb, and 4.75 kb in size, were completely contained within T-region, and each fragment of 9.7kb and 4.55 kb was joined to opposite ends of T-region. Also, it was identified that Sma I fragments which were contained within T-DNA region were covalently linked together within the pTi-12 DNA molecule.

참당귀(Angelica gigas Nakai) 뿌리의 면역증강활성 성분

안경섭,심웅섭,김환묵,한상배,김익환,Ahn, Kyung-Seop,Sim, Woong-Seop,Kim, Hwan-Mook,Han, Sang-Bae,Kim, Ik-Hwan 한국생약학회 1996 생약학회지 Vol.27 No.3

A potent immuno-stimulating activity was detected from the watersoluble and ethanol-precipitated crude extract (AG-1) of the root of Angelica gigas Nakai. The crude extract was fractionated into two fractions, an acidic AG-2 and a neutral AG-3 fraction by DEAE-cellulose adsorption. The two fractions contained polysaccharides of which M.W. were 10 Kdal and >2,000 Kdal respectively, proteins, and various inorganic components. The immunostimulating activities of two fractions were not reduced by proteinase K, acid or alkali treatment. The polysaccharides obtained from the root of A. gigas were mainly composed of arabinose, galactose, and galacturonic acid. These results indicated that immuno- stimulating components of the root of A. gigas was a kind of pectic polysaccharides or arabinogalactans.

Agrobacterium tumetaciens 및 Ti Plasmid DNA 에 의한 감자 ( Solanum tuberosum L . ) 괴경세포의 형질전환

정상호,심웅섭 ( Sang ho Chung,Woong Seop Sim ) 생화학분자생물학회 1987 BMB Reports Vol.20 No.4

In the present study, transformation of potato tuber cells by Agrobacterium tumefaciens and Ti plasmid treatment was studied. The results are as follows: Tumors were induced in potato tuber discs by infection with Agrobacterium tumefaciens KU-12. Whole plants were regenerated from the tumors on hormone-free medium. It was identified that the lysopine dehydrogenase (LpDH) activity was present in the tumor-derived plants. A total of 165 calli were obtained from Ti-plasmid DNA(pTi-12)-treated potato protoplasts on hormone-free medium and a few of them formed a root. Each of the non-rooted and rooted callus was randomly selected and tested for the presence of LpDH activity. Both of them were ocotopine positive, indicating that they were transformants. In order to analyze the T-DNA region maintained in transformed plant genome, the DNA-DNA hybridization experiment was performed. As a result, it was found that Sma I fragments of Ti plasmid (pTi-12) 13.4 kb, 3.7 kb, and 4.75 kb in size, were completely contained within T-region, and each fragment of 9.7kb and 4.55 kb was joined to opposite ends of T-region. Also, it was identified that Sma I fragments which were contained within T-DNA region were covalently linked together within the pTi-12 DNA molecule.

Chlorella 의 물질대사에 미치는 미양원소의 결핍효과(제 2 ) -, 리보 및 의 생합성능에 관하여-

이영록,진평,심웅섭,Lee, Yung-Nok,Chin, Pyung,Sim, Woong-Seop 한국미생물학회 1968 미생물학회지 Vol.6 No.1

Chlorella ellipsoidea cells were cultured in an iron, copper, zinc, manganese, molybdenum or boron-free medium. Biosynthetic activities of nucleic acids, protein and phospholipid in chlorella cells, which were growing in a microelement deficient medium were compared with those of the normal cells by measuring the contents of phosphate, amino acids or UV-absorbing substances in the various cell fractions. When the algae were grown in a molybdenum-free medium, the amounts of phosphate in the acid-soluble fraction of the cells increased, whereas the amounts of alkali-stable protein and RNA decreased compared with the normal cells showing that the synthesis of protein and RNA from the early products of photosynthesis was inhibited. When the algae were grown in a boron-free medium, amounts of alkali-labile protein and phospholipid of the cells decreased, while the amount of phosphate in acid-soluble fraction increased compared with the normal cells showing that the biosynthesis of protein and phospholipid from the early products of photosynthesis was retarded. In general, amounts of protein and RNA in the microelement deficient cells significantly decreased compared with those of the normal cells. Phosphate content in the acid-soluble fraction of the algal cell grown in an zinc, copper, molybdenum, or boron-free medium increased considerably, whereas that of the algal cell grown in an iron or manganese-free medium decreased remarkably compared with that of the control. It is considered, therefore, that molybdenum, zinc, copper and boron etc. play an important role in the biosyntbesis of macromolecule from acid-soluble phosphate compounds, in contrast to the principal action of iron and manganese on the photosynthetic process itself.

발아 중인 옥수수에서 리보조옴 단백질의 인산화반응에 미치는 GA3 의 효과

안경섭(Kyung Seop Ahn),음진성(Jin Seong Eum),심웅섭(Woong Seop Sim) 한국식물학회 1990 Journal of Plant Biology Vol.33 No.1

In order to study the effect of GA_3 on the phosphorylation of ribosomal proteins during germination in Zea mays, ribosomal proteins were labelled with ^32P, extracted, electrophoresed and autoradiographed. There are five phosphorylated ribosomal proteins. One of these is in 40S subunit and has molecular weight of 33,000 daltons. Others are in 60S subunit and have molecular weights of 37,000, 16,000, 15,200 and 13,500, respectively. Phosphorylation of ribosomal proteins was increased maximum 47.7% in shoots of Zea mays treated with GA_3.