• Title/Summary/Keyword: SUMO

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Transmission Performance Analysis of VCCN with SUMO depending on Packet Size and Participating Number of Clients (SUMO를 이용한 VCCN에서 패킷의 크기와 참여한 클라이언트 수에 따른 전송 성능에 대한 연구)

  • Yeon, Seunguk;Chae, Ye-Eun;Kang, Seung-Seok
    • The Journal of the Convergence on Culture Technology
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    • v.4 no.1
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    • pp.285-290
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    • 2018
  • Vehicular ad hoc network (VANET) is one of the future communication technologies in which it offers safe-driving information and Internet access for both drivers and passengers by communicating among cars on the road. In case of applying Content Centric Network (CCN) rather than TCP/IP over VANET, called VCCN, it can provide efficient multicast communication of shared information among participating cars as well as offer corresponding one-to-one communications. This paper introduces SUMO for simulating traffic flows, VANET for car-to-car ad hoc communications, and CCN for a future Internet architecture. In addition, it compares and analyses the transmission performance of moving cars over VCCN. According to the simulation results using SUMO and VCCN, the larger the packet size, the better the transmission performance. In addition, VCCN provides higher packet transmission rate than that of TCP/IP when the clients shares the same contents. Furthermore, the overall data reception rate exceeds the physical transmission channel rate.

SUMO Proteins are not Involved in TGF-${\beta}1$-induced, Smad3/4-mediated Germline ${\alpha}$ Transcription, but PIASy Suppresses it in CH12F3-2A B Cells

  • Lee, Sang-Hoon;Kim, Pyeung-Hyeun;Oh, Sang-Muk;Park, Jung-Hwan;Yoo, Yung-Choon;Lee, Junglim;Park, Seok-Rae
    • IMMUNE NETWORK
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    • v.14 no.6
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    • pp.321-327
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    • 2014
  • TGF-${\beta}$ induces IgA class switching by B cells. We previously reported that Smad3 and Smad4, pivotal TGF-${\beta}$ signal-transducing transcription factors, mediate germline (GL) ${\alpha}$ transcription induced by TGF-${\beta}1$, resulting in IgA switching by mouse B cells. Post-translational sumoylation of Smad3 and Smad4 regulates TGF-${\beta}$-induced transcriptional activation in certain cell types. In the present study, we investigated the effect of sumoylation on TGF-${\beta}1$-induced, Smad3/4-mediated $GL{\alpha}$ transcription and IgA switching by mouse B cell line, CH12F3-2A. Overexpression of small ubiquitin-like modifier (SUMO)-1, SUMO-2 or SUMO-3 did not affect TGF-${\beta}1$-induced, Smad3/4-mediated $GL{\alpha}$ promoter activity, expression of endogenous $GL{\alpha}$ transcripts, surface IgA expression, and IgA production. Next, we tested the effect of the E3 ligase PIASy on TGF-${\beta}1$-induced, Smad3/4-mediated $GL{\alpha}$ promoter activity. We found that PIASy overexpression suppresses the $GL{\alpha}$ promoter activity in cooperation with histone deacetylase 1. Taken together, these results suggest that SUMO itself does not affect regulation of $GL{\alpha}$ transcription and IgA switching induced by TGF-${\beta}1$/Smad3/4, while PIASy acts as a repressor.

Expression and Purification of Recombinant Human Epidermal Growth Factor Using Fusion Partners in Escherichia coli (융합 파트너를 이용한 인간 상피세포성장인자의 재조합 대장균에서 발현과 정제 연구)

  • Sung, Keehyun;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.56 no.5
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    • pp.711-717
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    • 2018
  • Human epidermal growth factor (hEGF) can stimulate the division of various cell types and has potential clinical applications. Since the protein contains three intra-molecular disulfide bonds, the high expression of active hEGF in Escherichia coli has not been well researched, We fused the hEGF gene with a small ubiquitin-related modifier gene (SUMO) by synthesizing an artificial SUMO-hEGF fusion gene that was highly expressed in E. coli (DE3) strain. The optimal expression level of the soluble fusion protein, SUMO-hEGF with IPTG (Isopropyl-${\beta}$-D-Thiogalactopyranoside), was up to 38.9% of the total cellular protein. The fusion protein was purified by Ni-NTA affinity chromatography and cleaved by a SUMO-specific protease to obtain the native hEGF, which was further purified by Ni-NTA affinity chromatography. The result of the reverse-phase HPLC showed that the purity of the recombinant cleaved hEGF was greater than 98%.

SUMO pathway is required for ribosome biogenesis

  • Hong-Yeoul, Ryu
    • BMB Reports
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    • v.55 no.11
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    • pp.535-540
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    • 2022
  • Ribosomes, acting as the cellular factories for protein production, are essential for all living organisms. Ribosomes are composed of both proteins and RNAs and are established through the coordination of several steps, including transcription, maturation of ribosomal RNA (rRNA), and assembly of ribosomal proteins. In particular, diverse factors required for ribosome biogenesis, such as transcription factors, small nucleolar RNA (snoRNA)-associated proteins, and assembly factors, are tightly regulated by various post-translational modifications. Among these modifications, small ubiquitin-related modifier (SUMO) targets lots of proteins required for gene expression of ribosomal proteins, rRNA, and snoRNAs, rRNA processing, and ribosome assembly. The tight control of SUMOylation affects functions and locations of substrates. This review summarizes current studies and recent progress of SUMOylation-mediated regulation of ribosome biogenesis.

Expression of Antimicrobial Peptide (AMP), Moricin Using SUMO Fusion Tag in Escherichia coli (대장균에서 SUMO fusion tag을 이용하여 항균펩타이드인 moricin의 발현)

  • Ahn, Dong-gyu;Park, Sun Ill;Kim, Soon Young
    • Journal of Life Science
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    • v.32 no.12
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    • pp.956-961
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    • 2022
  • Plant Chloroplast have several advantages as an expression platform of biopharmaceuticals over conventional expression platforms such as mammalian cells, yeast and bacteria. First, plants do not serve as a host for mammalian infectious virus and have endotoxin like bacteria which can cause anaphylactic shock. In addition, high copy number of chloroplast genome allows for chloroplast transformants to reach the high level of expression of heterologous genes. Moreover, the integration of transgenes into specific region of chloroplast genomes makes chloroplast transformants unaffected by positional effect which can be frequently observed from nuclear transformants, resulting in loss of transgene expressions. Antimicrobial peptides (AMPs) are a kind of innate immunity which is found from bacteria to humans. Unlike conventional antibiotics, very less dosage of AMPs can have catastrophic effect on bacterial survival. Further, the repeated use of AMPs does not trigger the development of bacterial resistance. Moricin, one of the AMPs, was isolated from Bombyx mori, a silkworm moth. The C-terminal of moricin consists largely of basic amino acids, and the N-terminal has an α-helix structure. Moricin was chosen and expressed in a SUMO/SUMOase without leaving any unwanted amino acids which could potentially affect the anti-bacterial activity of the moricin. The transformation vector used in this study has already been created in this lab for the expression in both prokaryotic systems such as E. coli and chloroplast. The expressed moricin was purified using Ni columns and SUMOase, and the antibacterial activity of the purified moricin was confirmed using an agar diffusion assay.

Host Cell Nuclear Localization of Shigella flexneri Effector OspF Is Facilitated by SUMOylation

  • Jo, Kyungmin;Kim, Eun Jin;Yu, Hyun Jin;Yun, Cheol-Heui;Kim, Dong Wook
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.610-615
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    • 2017
  • When Shigella infect host cells, various effecter molecules are delivered into the cytoplasm of the host cell through the type III secretion system (TTSS) to facilitate their invasion process and control the host immune responses. Among these effectors, the S. flexneri effector OspF dephosphorylates mitogen-activated protein kinases and translocates itself to the nucleus, thus preventing histone H3 modification to regulate expression of proinflammatory cytokines. Despite the critical role of OspF, the mechanism by which it localizes in the nucleus has remained to be elucidated. In the present study, we identified a potential small ubiquitin-related modifier (SUMO) modification site within OspF and we demonstrated that Shigella TTSS effector OspF is conjugated with SUMO in the host cell and this modification mediates the nuclear translocation of OspF. Our results show a bacterial virulence factor can exploit host post-translational machinery to execute its intracellular trafficking.

Identification of Chinese Cabbage Sentrin as a Suppressor of Bax-Induced Cell Death in Yeast

  • Sawitri, Widhi Dyah;Slameto, Slameto;Sugiharto, Bambang;Kim, Kyung-Min
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.600-606
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    • 2012
  • Studies into the cell death program termed apoptosis have resulted in new information regarding how cells control and execute their own demise, including insights into the mechanism by which death-preventing factors can inhibit Bax-induced caspase activation. We investigated high temperature stress-induced cell death in Brassica rapa. Using a yeast functional screening from a Brassica rapa cDNA library, the BH5-127 EST clone encoding an apoptotic suppressor peptide was identified. However, a phylogenic tree showed that BH5-127 clusters within a clade containing SUMO-1 (Small Ubiquitin-like Modifier-1). BH5-127 was confirmed similar to have function to SUMO-1 as Fas suppression. Expression of BH5-127 showed that substantial suppression of cell death survived on SD-galactose-$Leu^-$-$Ura^-$ medium. The results suggest that BrSE ($\underline{B}$rassica rapa $\underline{S}$entrin $\underline{E}$ST, BH5-127) is one of the important regulatory proteins in programming cell death, especially in the seedling stage of Chinese cabbage.

Analysis of a Region Required for the Functions of Fission Yeast Nucleoporin Nup184 and Its SUMO Modification (분열효모 핵공단백질인 Nup184의 기능에 필요한 부위 분석 및 SUMO 변성)

  • Chai, Ai-Ree;Jang, Soo-Yeon;Yoon, Jin-Ho
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.66-72
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    • 2012
  • The Nup188 protein is one of the largest evolutionally conserved nucleoprins (Nups) that compose the inner ring of nuclear pore complex (NPC). The Nup184 protein, fission yeast Schizosaccharomyces pombe ortholog of Nup188p, is required for normal growth and mRNA export in nutrient-rich medium (YES). Here, we identified a carboxyl region (482 to 1628) of Nup184 protein that was enough to complement the defects of both growth and mRNA export when the ${\Delta}nup184$ knock-out mutant was grown in YES medium. This region is also required for localization of GFP-Nup184 fusion to the nuclear periphery. In addition, we found that ORF of Nup184 (predicted 1564 amino-acid protein) registered in S. pombe GeneDB (hosted by Sanger Institute, UK) is 64 amino-acid residues shorter than that predicted by our sequence data. This carboxy-terminal region is necessary for the functions of Nup184p. We further demonstrated that Nup184 protein was conjugated with SUMO in vivo.