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Subcellular Localization of the Barley Stripe Mosaic Virus Triple Gene Block Proteins
Lim, Hyoun-Sub,Bragg, Jennifer N.,Ganesan, Uma,Ruzin, Steven,Schichnes, Denise,Lee, Mi Yeon,Vaira, Anna Maria,Ryu, Ki Hyun,Hammond, John,Jackson, Andrew O. American Society for Microbiology 2009 Journal of virology Vol.83 No.18
<B>ABSTRACT</B><P><I>Barley stripe mosaic virus</I> (BSMV) spreads from cell to cell through the coordinated actions of three triple gene block (TGB) proteins (TGB1, TGB2, and TGB3) arranged in overlapping open reading frames (ORFs). Our previous studies (D. M. Lawrence and A. O. Jackson, J. Virol. 75:8712-8723, 2001; D. M. Lawrence and A. O. Jackson, Mol. Plant Pathol. 2:65-75, 2001) have shown that each of these proteins is required for cell-to-cell movement in monocot and dicot hosts. We recently found (H.-S. Lim, J. N. Bragg, U. Ganesan, D. M. Lawrence, J. Yu, M. Isogai, J. Hammond, and A. O. Jackson, J. Virol. 82:4991-5006, 2008) that TGB1 engages in homologous interactions leading to the formation of a ribonucleoprotein complex containing viral genomic and messenger RNAs, and we have also demonstrated that TGB3 functions in heterologous interactions with TGB1 and TGB2. We have now used <I>Agrobacterium tumefaciens</I>-mediated protein expression in <I>Nicotiana benthamiana</I> leaf cells and site-specific mutagenesis to determine how TGB protein interactions influence their subcellular localization and virus spread. Confocal microscopy revealed that the TGB3 protein localizes at the cell wall (CW) in close association with plasmodesmata and that the deletion or mutagenesis of a single amino acid at the immediate C terminus can affect CW targeting. TGB3 also directed the localization of TGB2 from the endoplasmic reticulum to the CW, and this targeting was shown to be dependent on interactions between the TGB2 and TGB3 proteins. The optimal localization of the TGB1 protein at the CW also required TGB2 and TGB3 interactions, but in this context, site-specific TGB1 helicase motif mutants varied in their localization patterns. The results suggest that the ability of TGB1 to engage in homologous binding interactions is not essential for targeting to the CW. However, the relative expression levels of TGB2 and TGB3 influenced the cytosolic and CW distributions of TGB1 and TGB2. Moreover, in both cases, localization at the CW was optimal at the 10:1 TGB2-to-TGB3 ratios occurring in virus infections, and mutations reducing CW localization had corresponding effects on BSMV movement phenotypes. These data support a model whereby TGB protein interactions function in the subcellular targeting of movement protein complexes and the ability of BSMV to move from cell to cell.</P>
대한산업보건협회 산업보건연구소의 미국 정도관리 참여결과
이선주,최호춘,박찬우,오미순,이준승,전수경,박화미,정규철,Lee, Seon-Ju,Choe, Ho-Chun,Park, Chan-U,O, Mi-Sun,Lee, Jun-Seung,Jeon, Su-Gyeong,Park, Hwa-Mi,Jeong, Gyu-Cheol 대한산업보건협회 1995 산업보건 Vol.88 No.-
PAT program(The Proficiency Analytical Testing Program)은 미국산업위생협회(American Industrial Hygiene Association AIHA)와 미국산업안전보건연구원(National Institute for Occupational Safety and Health NIOSH)에서 공동주관하는 정도관리로서 일년에 4회 실시하고 있다. 실시항목은 금속(납, 카드뮴, 크롬, 아연), 유리규산, 석면, 유기,용제(Benzen, O-Xylene, Toluene, Carbon Tetrachloride, 1, 1, 1-Trichloroethane, Chloroform, Tetrachloroethylene, 1, 2-Dichloroethane, methanol, p-Dioxane)이다. 대한산업보건협회 산업보건연구소에서는 위의 4가지 항목을 모두 신청하여 1995년 1월에 실시된 120회 부터 참여하였는데 다음은 본 연구소의 120화에 대한 분석방법과 NIOSH로 부터 받은 결과이다.
갑상선 세포에서 Peroxiredoxin에 의한 과산화수소 및 아포프토시스 조절
김현진,이태훈,김도희,권오유,김영건,송민호,노흥규,박수정,김호,박은신,정효균,서재미,채수홍 대한내분비학회 2000 Endocrinology and metabolism Vol.15 No.1
Background : Peroxiredoxins(Prx) play an important role in regulating cellular differentiation and proliferation in several types of mammalian cells. One mechanism for this action involves modulation of hydrogen peroxide(H2O2)-mediated cellular responses. This report examines the expression of Prx I and Prx II in thyroid cells and their roles in eliminating H2O2 produced in response to TSH. Methods : The expression of Prx-I and Prx-II were quantiated in FRTL-5 after stimulation with Thyroid stimulating hormone (TSH), Forskolin (FSK), Methimazole (MMI) and hydrogen peroxide (H2O2). Transient transfections were carried out with FRTL-5 cells at 80% confluency and 20?g of pCRprx I and pCRprx II or equivalent molar amounts of the pCR3.1TM basic vector. Transient transfection used an electroporation technique. Intracellular H2O2 was assayed in FRTL-5 cells with a fluorescent dye, 2', 7'-dichlorofluoresceindiacetate(DCFH-DA). Apoptosis of cells were evaluated by using an detection kit (Promega, Inc., Madison, WI). Results : Prx I and Prx II are constitutively expressed in FRTL-5 thyroid cells. Prx I expression, but not Prx II expression, is stimulated by exposure to TSH and H2O2. In addition, methimazole(MMI) induces a high level of Prx I mRNA and protein in these cells. Overexpression of Prx I and Prx II enhance the elimination of H2O2 produced by TSH in FRTL-5 cells. Treatment with 500?M H2O2 causes apoptosis in FRTL-5 cells as evidenced by standard assays of apoptosis(i.e., terminal deoxynucleotidyl transferase deoxyuridine triphosphate-biotin nick end-labeling(TUNEL), BAX expression and PARP cleavage. Overexpression of Prx I and Prx II reduces the amount of H2O2-induced apoptosis measured by these assays. Conclusion : These results suggest that Prx I and Prx II are involved in the removal of H2O2 in thyroid cells, and can protect these cells from undergoing apoptosis. These proteins are likely to be involved in the normal physiological response to TSH-induced production of H2O2 in thyroid cells(J Kor Soc Endocrinol15:55-69, 2000).
USP14 inhibition regulates tumorigenesis by inducing apoptosis in gastric cancer
Mi Yea Lee,Min-Jee Kim,Jun-O Jin,Peter Chang-Whan Lee 생화학분자생물학회 2023 BMB Reports Vol.56 No.8
Deubiquitinases (DUBs) are an essential component of theubiquitin-proteasome system (UPS). They trim ubiquitin fromsubstrate proteins, thereby preventing them from degradation,and modulate different cellular processes. Ubiquitin-specificprotease 14 (USP14) is a DUB that has mainly been studied forits role in tumorigenesis in several cancers. In the present study,we found that the protein levels of USP14 were remarkablyhigher in gastric cancer tissues than in the adjacent normaltissues. We also demonstrated that the inhibition of USP14activity using IU1 (an USP14 inhibitor) or the inhibition ofUSP14 expression using USP14-specific siRNA markedly reducedthe viability of gastric cancer cells and suppressed their migratoryand invasive abilities. The reduction in gastric cancercell proliferation due to the inhibition of USP14 activity was aresult of the increase in the degree of apoptosis, as evidencedby the increased expression levels of cleaved caspase-3 andcleaved PARP. Furthermore, an experiment using the USP14inhibitor IU1 revealed that the inhibition of USP14 activitysuppressed 5-fluorouracil (5-FU) resistance in GC cells. Collectively,these findings indicate that USP14 plays critical roles ingastric cancer progression and suggest its potential to serve asa novel therapeutic target for gastric cancer treatment.
Subspecific Status of the Korean Tiger Inferred by Ancient DNA Analysis
Lee, Mu-Yeong,Hyun, Jee-Yun,Lee, Seo-Jin,An, Jung-Hwa,Lee, Eun-Ok,Min, Mi-Sook,Kimura, Junpei,Kawada, Shin-Ichiro,Kurihara, Nozomi,Luo, Shu-Jin,O'Brien, Stephen J.,Johnson, Warren E.,Lee, Hang The Korean Society of Systematic Zoology 2012 Animal Systematics, Evolution and Diversity Vol.28 No.1
The tiger population that once inhabited the Korean peninsula was initially considered a unique subspecies (Panthera tigris coreensis), distinct from the Amur tiger of the Russian Far East (P. t. altaica). However, in the following decades, the population of P. t. coreensis was classified as P. t. altaica and hence forth the two populations have been considered the same subspecies. From an ecological point of view, the classification of the Korean tiger population as P. t. altaica is a plausible conclusion. Historically, there were no major dispersal barriers between the Korean peninsula and the habitat of Amur tigers in Far Eastern Russia and northeastern China that might prevent gene flow, especially for a large carnivore with long-distance dispersal abilities. However, there has yet to be a genetic study to confirm the subspecific status of the Korean tiger. Bone samples from four tigers originally caught in the Korean peninsula were collected from two museums in Japan and the United States. Eight mitochondrial gene fragments were sequenced and compared to previously published tiger subspecies' mtDNA sequences to assess the phylogenetic relationship of the Korean tiger. Three individuals shared an identical haplotype with the Amur tigers. One specimen grouped with Malayan tigers, perhaps due to misidentification or mislabeling of the sample. Our results support the conclusion that the Korean tiger should be classified as P. t. altaica, which has important implications for the conservation and reintroduction of Korean tigers.
Analysis of genetic diversity and cytoplasm male-sterility types in radish germplasm
O New Lee,Hyo Joung Kwon,Mi Kyung Han,Han Yong Park 한국육종학회 2015 한국육종학회 심포지엄 Vol.2015 No.07
Radish (Raphanus sativus L.) is a widely-consumed root vegetable that is grown worldwide. To utilize the radish genetic resources for breeding research, we collected radish germplasms and evaluated their morphological and genetical characteristics. Here, phylogenetic relationship of 288 accessions were analyzed using 16 SSR markers and classified cytoplasm male sterility (CMS) types using cpDNA-based molecular markers. To create a collection of 288 accessions, 188 and 73 accessions were selected from RDA-Genebank (Korea) and NIAS-Genebank (Japan), respectively, after generation advancement for the accessions with low uniformity. In addition, 27 elite lines currently used for commercial radish breeding programs were included. In the result of phylogenetic analysis, 288 accessions were clustered into 5 major groups corresponding to the morphological traits and origins at the similarity coefficient value of 0.51. Analysis of CMS types revealed that majority of accessions were determined as DBRMF1 and DBRMF2 mitotypes, 15 accessions to Ogura and 4 accessions to DCGMS mitotypes. Further genetic analysis for radish germplasm will be valuable in assisting radish f1 hybrid breeding.