• Journal of Microbiology and Biotechnology
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• 제15권3호
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• pp.484-490
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• 2005

Human ribosomal protein S3 (rpS3), which has a DNA repair endonuclease activity, is a multifunctional protein. This protein is involved in DNA repair, translation, and apoptosis. In particular, rpS3 has a lyase activity, which cleaves the phosphodiester bond of damaged sites such as cyclobutane pyrimidine dimers and AP sites. Here, using deletion analysis, we identified that the repair endonuclease domain resides in the C-terminal region (165-243 aa) of rpS3. We also found that ectopic expression of GST-rpS3 in bacterial strain BL21 promoted the resistance of these cells to ultraviolet (UV) radiation and hydrogen peroxide ($H_{2}O_{2}$) treatment. The repair domain of rpS3 was sufficient to exhibit the resistance to UV irradiation and recover cell growth and viability, showing that the repair activity of rpS3 is responsible for the resistance to UV irradiation. Our study suggests that rpS3 is able to process DNA damage in bacteria via its repair domain, showing the resistance to genotoxic stress. This implies that rpS3-like activity could be operative in bacteria.

• BMB Reports
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• 제56권5호
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• pp.302-307
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• 2023

Lyn, a tyrosine kinase that is activated by double-stranded DNA-damaging agents, is involved in various signaling pathways, such as proliferation, apoptosis, and DNA repair. Ribosomal protein S3 (RpS3) is involved in protein biosynthesis as a component of the ribosome complex and possesses endonuclease activity to repair damaged DNA. Herein, we demonstrated that rpS3 and Lyn interact with each other, and the phosphorylation of rpS3 by Lyn, causing ribosome heterogeneity, upregulates the translation of p-glycoprotein, which is a gene product of multidrug resistance gene 1. In addition, we found that two different regions of the rpS3 protein are associated with the SH1 and SH3 domains of Lyn. An in vitro immunocomplex kinase assay indicated that the rpS3 protein acts as a substrate for Lyn, which phosphorylates the Y167 residue of rpS3. Furthermore, by adding various kinase inhibitors, we confirmed that the phosphorylation status of rpS3 was regulated by both Lyn and doxorubicin, and the phosphorylation of rpS3 by Lyn increased drug resistance in cells by upregulating p-glycoprotein translation.

• BMB Reports
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• 제44권8호
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• pp.529-534
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• 2011

Ribosomal protein S3 (rpS3) is a multifunctional protein involved in translation, DNA repair, and apoptosis. The relationship between rpS3 and cyclin-dependent kinases (Cdks) involved in cell cycle regulation is not yet known. Here, we show that rpS3 is phosphorylated by Cdk1 in G2/M phase. Co-immunoprecipitation and GST pull-down assays revealed that Cdk1 interacted with rpS3. An in vitro kinase assay showed that Cdk1 phosphorylated rpS3 protein. Phosphorylation of rpS3 increased in nocodazole-arrested mitotic cells; however, treatment with Cdk1 inhibitor or Cdk1 siRNA significantly attenuated this phosphorylation event. The phosphorylation of a mutant form of rpS3, T221A, was significantly reduced compared with wild-type rpS3. Decreased phosphorylation and nuclear accumulation of T221A was much more pronounced in G2/M phase. These results suggest that the phosphorylation of rpS3 by Cdk1 occurs at Thr221 during G2/M phase and, moreover, that this event is important for nuclear accumulation of rpS3.

• Journal of Photoscience
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• 제10권2호
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• pp.195-198
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• 2003

In the cellular response to DNA damaging agents, cells undergo cell cycle arrest or apoptosis against irrepairable DNA damage. RpS3 is known to function as UV DNA repair endonuclease III and ribosomal protein S3. In this study, we used normal and rpS3-overexpressed 293T cells to examine the role of rpS3 in response to DNA damaging agents. When 293T cells transfected with rpS3 were irradiated with UV, the pattern of cell cycle was dramatically changed in comparison with un-transfected 293T cells. We also found that the expression of rpS3 in normal cells was increased by treatment with DNA damaging agents. By means of Western and Northern blot analyses in rat tissues, we showed the expression pattern of rpS3 protein and its mRNA. These data suggest that DNA repair and cell cycle arrest are interrelated to each other through rpS3, and the increased expression of rpS3 seems to regulate the cell cycle arrest by DNA damaging agents.

• BMB Reports
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• 제39권2호
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• pp.208-215
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• 2006

The human ribosomal protein S3 (rpS3) was expressed in E. coli using the pET-I5b vector and the monoclonal antibodies (mAbs) were produced and characterized. A total of five hybridoma cell lines were established and the antibodies recognized a single band of molecular weight of 33 kDa on immunoblot with purified rpS3. When the purified rpS3 was incubated with the mAbs, the UV endonuclease activity of rpS3 was inhibited up to a maximum of 49%. The binding affinity of mAbs to rpS3 determined by using a biosensor technology showed that they have similar binding affinities. Using the anti-rpS3 antibodies as probes, we investigated the cross-reactivities of various other mammalian brain tissues and cell lines, including human. The immunoreactive bands on Western blots appeared to be the same molecular mass of 33 kDa in all animal species tested. They also appear to be extensively cross-reactive among different organs in rat. These results demonstrated that only one type of immunologically similar rpS3 protein is present in all of the mammalian brain tissues including human. Furthermore, these antibodies were successfully applied in immunohistochemistry in order to detect rpS3 in the gerbil brain tissues. Among the various regions in the brain tissues, the rpS3 positive neurons were predominantly observed in the ependymal cells, hippocampus and substantia nigra pars compacta. The different distributions of rpS3 in brain tissues reply that rpS3 protein may play an important second function in the neuronal cells.

• 미생물학회지
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• 제26권3호
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• pp.173-180
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• 1988

염색체 유전자를 전달시키기 위한 기초작업으로 RP4 :: Mu cts와 RP4 :: mini-Mu 잡종 플라스미드를 접합에 의하여 E.. coli로 부터 Pseudomonas로 전달시켰다. RP4::Mu cts와 RP4:: mini-Mu의 수용세포는 플라스미드를 가지지 아니하는 Pseudomonas 균주들의 항생세 내성, 탄화수소 자화능 등의 유전적 지표를 조사하여 사용하였다. RP4::mini-Mu는 1$10^{-2}$ - $10^{-4}$의 빈도로 전달되었으며 RP4:: Mu cts는 Pseµdomonas aeruginosa KU557로는 $10^{-2}$의 빈도로, 그 이외의 수용세포로는 10?7의 빈도로 천달되었다. 접합체에 전달된 플라스미드의 존재는 암피실린, 카나마이신, 테트라싸이클린에 대한 내성과 전기영돔에 의해 확인하였으며 특히 RP4::Mu cts는 $43^{\circ}C$에서의 플라크 행성으로도 확인하였다. 접합체들로 부터 생성된 Mu 파아지는 약 $10^{5}$의 P.F.U.를 나타냈으며 전달된 RP4:: Mu cts와 RP4:: mini-Mu는 접합체들에서 비교적 안장한 것으로 밝혀졌다.

• 한국CDE학회지
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• 제2권2호
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• pp.12-16
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• 1996

Rapid Prototyping(RP)이란 짧은 시간 내에 CAD 그래픽 데이터로부터 3차원 형상의 시제품을 만들어 내는 기술을 일컬으며, 1986년 3D Systems라는 회사에 의하여 상용화된 SLA(Stereolithography Apparatus)방식을 선두로 하여 지난 10년간 급속히 발전을 해왔다. 원하는 형상의 부품을 만들 때 기존의 machining은 원자재에서 재료를 깎아내면서 만드는 반면에, RP는 재료를 한 층씩 차례로 쌓아서 부품을 만드는 가산적인 공정 특징을 가지고 있다. 액체, 고체, 심지어 기체 상태의 재료까지 다양한 재료를 사용하고 있으며 또한 여러가지 적층방법으로 부품을 제작하고 있다. 성공적인 RP기술의 창출에는 RP기계제작에 직접 관계되는 기술뿐만 아니라, 재료 기술, RP제작에 적합한 CAD데이터 생성기술, 후처리 및 가공기술 등이 모두 관건이 된다. 여기서는 RP기술의 주요 파트 제작방법과 RP에 쓰이는 재료, RP의 용도 및 그 한계성 등에 대하여 생각하여 보았다. RP기술은 3차원 CAD 모델이 없으면 실현이 불가능하다. 3차원(3D)화는 제품을 설계하고 만드는 대부분의 회사가 경쟁력을 갖기 위하여 싫든 좋든 이루어야 하는 목표 중의 하나라고 할 수 있는데, RP기술 도입은 이러한 3차원화를 단축시키는 촉매제의 역할을 할 수 있다고 생각한다. RP기술이 부분적으로 정확도의 문제와 제작 가능한 재료의 종류 및 성능에 제한이 있지만, 현재로서도 여러 응용분야에 성공적으로 이용되고 있으며 향후에는 더욱 그 응용 범위를 넓혀갈 것으로 전망된다.

• Journal of Photoscience
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• 제9권2호
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• pp.182-185
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• 2002

There are 3 UV DNA repair endonuclease activities in mammalian cells that cleave UV -irradiated DNA. Interestingly, mammalian UV endonuclease III with MW of 26.7kD has a lyase activity on AP sites. It also cleaves the phosphodiester bond within a cyclobutane pyrimidine dimer. Genomic analysis of human repair endonuclease III gene revealed that this gene has 100% sequence identity with ribosomal protein S3 (rpS3). Therefore, rpS3 seems to function both in translation and in DNA repair. This gene of about 6.1 kb contains 6 introns and 7 exons, and the first and fifth introns of human rpS3 gene contain functional U15 small nucleolar (sno) RNAs which appear to be involved in ribosome assembly. It is to be noted that the column profile of the endonuclease activity of rpS3 appears to be altered in Xeroderma Pigmentosum (XP) group D cells compared to normal cells indicating that this protein is involved in XP disease as well. XP is a human disease characterized by high sensitivity of skin by UV- or sun-light irradiation and by high frequency of developing skin cancers. We also report here that rpS3 protein is involved in other cellular functions.

• Journal of Microbiology
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• 제38권2호
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• pp.88-92
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• 2000

A human ribosomal protein S3 (rpS3), which also functions as a DNA repair enzyme(UV endonuclease III), was expressed in a methylotrophic yeast, Pichia pastoris, and biochemically characterized. UV endonuclease activity was preiously characterized, and this activity of mammalian rpS3 was found to be non-specfic upon purification and storage. Under the Pichia expression system, the subcloned cDNA of the human rpS3 gene revealed a peptide of 42 kDa by SDS-PAGE and Western blot. The secreted form of human rpS3 rendered no endonuclease activity while the intracellular form showed UV specific endonuclease activity by the nick circle assay.

• 미생물학회지
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• 제23권2호
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• pp.107-114
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• 1985

우리나라 자연환경으로부터 분리한 Rhizob ium 32권주로부터 카나마이신, 세트라싸이클린 등에는 예민하고, 클로람페니콜, 젠타마이신 등에는 강한 내성을 나타내는 5 균주를 선별하고, 이를 수용세포로하여 RP4::Mu cts를 접함에 의해 E. coli로부터 Rhizobium leguminosarum으로 전달 시켰다. 그 전달빈도는 $5.8{\times}10^{-7}$의 빈도를 나타내었다. 접합체에서의 RP4::Mu cts플라스미드의 존재는 암피실린과 카나마이신, 테트라싸이클린에 대한 내정과 $42^{\circ}C$에서의 플라크 형성으로 확인하였다. 접합체들은 $10^2~10^3$단위로 플라크를 형성하였고, 안정성를 조사한 신파 4주 후에도 대부분이 RP4: :Mu cts의 성질을 유지하고 있는 것으로 나타났다.