• Title/Summary/Keyword: RNA transcript

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Expression of Co-stimulatory Molecules and STAT/SOCS Signaling Factors in the Splenocytes of Mice Tolerized against Arthritis by Oral Administration of Type II Collagen (제2형 콜라겐으로 경구관용을 유도한 관절염 모델 마우스의 비장림프구내의 보조자극인자 및 STAT/SOCS 신호전달 인자의 발현 양상조사)

  • Lee, Kang-Eun;Hwang, Sue-Yun;Min, So-Youn;Kim, Ho-Youn
    • IMMUNE NETWORK
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    • v.3 no.3
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    • pp.248-254
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    • 2003
  • Oral administration of antigen has long been used in the induction of immune tolerance in various animal models of autoimmune diseases including rheumatoid arthritis (RA). Alleveation of arthritogenic symptoms has been reported from RA patients who received oral administration of type II collagen (CII) without side effects, however its rather inconsistent therapeutic efficacy and variation among patients calls for more detailed investigation on the mechanism of oral tolerance to be settled as regular treatment for RA. In an attempt to understand the immunogenic processes underpinning tolerance induction by orally administered CII, we analyzed changes in the expression of costimulatory molecules and STAT/SOCS signaling messengers in the mouse model of collagen induced arthritis (CIA). We found thatin the spleen of CIA mice, that has been undergone repeated oral feeding of CII prior to the induction of arthritis, showed increased promortion of CTLA4 expressing lymphocytes than in the spleen of PBS fed control. On the other hand, cells expressing CD28 or ICOS were decreased in the spleen of tolerized mice. Tolerance induction by oral CII administration also enhanced the expression of STAT6 in both RNA and protein level, while not affecting the expression of STAT3. The expression of SOCS3, which hasbeen known to transmit STAT-mediated signals from Th2 type cytokines, remained unchanged in the spleen of tolerized mice. Interestingly transcript of SOCS1, which has been associated with Th1 related pathways, was only visible in the spleen of tolerized but not of control mice, suggesting that as in the case of IL-6 signaling, it may exert a feed back inhibition toward the Th1 type stimulation.

Structure and Expression Analyses of SVA Elements in Relation to Functional Genes

  • Kwon, Yun-Jeong;Choi, Yuri;Eo, Jungwoo;Noh, Yu-Na;Gim, Jeong-An;Jung, Yi-Deun;Lee, Ja-Rang;Kim, Heui-Soo
    • Genomics & Informatics
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    • v.11 no.3
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    • pp.142-148
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    • 2013
  • SINE-VNTR-Alu (SVA) elements are present in hominoid primates and are divided into 6 subfamilies (SVA-A to SVA-F) and active in the human population. Using a bioinformatic tool, 22 SVA element-associated genes are identified in the human genome. In an analysis of genomic structure, SVA elements are detected in the 5′ untranslated region (UTR) of HGSNAT (SVA-B), MRGPRX3 (SVA-D), HYAL1 (SVA-F), TCHH (SVA-F), and ATXN2L (SVA-F) genes, while some elements are observed in the 3′UTR of SPICE1 (SVA-B), TDRKH (SVA-C), GOSR1 (SVA-D), BBS5 (SVA-D), NEK5 (SVA-D), ABHD2 (SVA-F), C1QTNF7 (SVA-F), ORC6L (SVA-F), TMEM69 (SVA-F), and CCDC137 (SVA-F) genes. They could contribute to exon extension or supplying poly A signals. LEPR (SVA-C), ALOX5 (SVA-D), PDS5B (SVA-D), and ABCA10 (SVA-F) genes also showed alternative transcripts by SVA exonization events. Dominant expression of HYAL1_SVA appeared in lung tissues, while HYAL1_noSVA showed ubiquitous expression in various human tissues. Expression of both transcripts (TDRKH_SVA and TDRKH_noSVA) of the TDRKH gene appeared to be ubiquitous. Taken together, these data suggest that SVA elements cause transcript isoforms that contribute to modulation of gene regulation in various human tissues.

Expression of a Gene Encoding Heat shock Protein 70-Related Protein from Olive Flounder, Paralichthys olivaceus

  • Kim, Woo-Jin;Lee, Jeong-Ho;Kim, Kyung-Kil;Park, Jung-Youn;Kang, Ho-Sung;Kim, Han-Do
    • Journal of Aquaculture
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    • v.12 no.3
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    • pp.175-183
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    • 1999
  • We have shown previously that the sequence of olive flounder (Paralichthys olivaceus) hsp70-related cDAN has a high similarity with those of cognate hsc70 of other species (Kim et al., 1999; J. Aquaculture, 12:91-100). In order to investigate whether this gene encodes the congate hsc70, we examined the expression of this gene in normal and heat-shocked conditions. By in vitro translation, this gene encoded a 70 kD protein which was constitutively experessed and was not induced by heat shock. This translated protein was recognized by anti-hsp/hsc70 antibody. Tests of heat-inducibility showed that this gene was constitutively expressed in normal conditions and its expression was not increased after heat shock. The expression levels of this gene were high in stomach, gill, intestine, kidney and brain, moderate in liver, and comparatively low in overy and heart. Furthermore, Northern blot analysis of transcript expression showed that the corresponding mRNA were detected throughout embryonic development in the absence of any heat shock. These results provided evidence that olive flounder hsp70-related cDNA encoded to cognate member of hsp70 family, hsc70.

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Expression and Characterization of ${\alpha}$-Methylacyl CoA Racemase from Anisakis simplex Larvae

  • Kim, Bong-Jin;Kim, Sun-Mi;Cho, Min-Kyung;Yu, Hak-Sun;Lee, Yong-Seok;Cha, Hee-Jae;Ock, Mee-Sun
    • Parasites, Hosts and Diseases
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    • v.50 no.2
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    • pp.165-171
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    • 2012
  • Larval excretory-secretory products of Anisakis simplex are known to cause allergic reactions in humans. A cDNA library of A. simplex 3rd-stage larvae (L3) was immunoscreened with polyclonal rabbit serum raised against A. simplex L3 excretory-secretory products to identify an antigen that elicits the immune response. One cDNA clone, designated as ${\alpha}$-methylacyl CoA racemase (Amacr) contained a 1,412 bp cDNA transcript with a single open reading frame that encoded 418 amino acids. A. simplex Amacr showed a high degree of homology compared to Amacr orthologs from other species. Amacr mRNA was highly and constitutively expressed regardless of temperature (10-$40^{\circ}C$) and time (24-48 hr). Immunohistochemical analysis revealed that Amacr was expressed mainly in the ventriculus of A. simplex larvae. The Amacr protein produced in large quantities from the ventriculus is probably responsible for many functions in the development and growth of A. simplex larvae.

Inhibition of MicroRNA-221 and 222 Enhances Hematopoietic Differentiation from Human Pluripotent Stem Cells via c-KIT Upregulation

  • Lee, Ji Yoon;Kim, MyungJoo;Heo, Hye-Ryeon;Ha, Kwon-Soo;Han, Eun-Taek;Park, Won Sun;Yang, Se-Ran;Hong, Seok-Ho
    • Molecules and Cells
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    • v.41 no.11
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    • pp.971-978
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    • 2018
  • The stem cell factor (SCF)/c-KIT axis plays an important role in the hematopoietic differentiation of human pluripotent stem cells (hPSCs), but its regulatory mechanisms involving microRNAs (miRs) are not fully elucidated. Here, we demonstrated that supplementation with SCF increases the hematopoietic differentiation of hPSCs via the interaction with its receptor tyrosine kinase c-KIT, which is modulated by miR-221 and miR-222. c-KIT is comparably expressed in undifferentiated human embryonic and induced pluripotent stem cells. The inhibition of SCF signaling via treatment with a c-KIT antagonist (imatinib) during hPSC-derived hematopoiesis resulted in reductions in the yield and multi-lineage potential of hematopoietic progenitors. We found that the transcript levels of miR-221 and miR-222 targeting c-KIT were significantly lower in the pluripotent state than they were in terminally differentiated somatic cells. Furthermore, suppression of miR-221 and miR-222 in undifferentiated hPSC cultures induced more hematopoiesis by increasing c-KIT expression. Collectively, our data implied that the modulation of c-KIT by miRs may provide further potential strategies to expedite the generation of functional blood cells for therapeutic approaches and the study of the cellular machinery related to hematologic malignant diseases such as leukemia.

Identification of genes related to ER stress in bZIP28 gene transgenic potato plant

  • Kim, Dool Yi;Kim, Kyung Hwa;Choi, Man Soo;Ok, Hyun Choog;Kim, Jae Hyun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.153-153
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    • 2017
  • Potato (Solanum tubersosum L.) is susceptible to various environmental stresses such as frost, high temperature, and drought. Enhancement of potato drought tolerance can reduce yield loss under drought that has negative effect on potato tuber growth. Genetic engineering can be utilized to achieve this goal, but such approaches using endogenous potato genes have rarely been applied. Since unpredictable global weather changes cause more severe and frequent water limiting conditions, improvement of potato drought tolerance can minimize such adverse effects under drought and can impact on sustainable potato production. Genetic engineering can be utilized to improve potato drought tolerance, but such approaches using endogenous potato genes have rarely been applied. We were obtained AtbZIP28 gene transgenic potato plants. It is identified transcript levels at various stress conditions, polyethylene glycol (PEG), NaCl, abscisic ${\underline{acid}}$ (ABA). Also, For identification to regulate ER stress response genes in AtbZIP28 gene transgenic potato plant, we screened seven potato genes from RNA-seq analysis under TM treatment. Five and two genes were up- and down-regulated by TM, respectively. Their expression patterns were re-examined at stress agents known to elicit TM, DTT, DMSO and salt stress.

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Elicitation of Penicillin Biosynthesis by Alginate in Penicillium chrysogenum, Exerted on pcbAB, pcbC, and penDE Genes at the Transcriptional Level

  • Liu, Gang;Casqueiro, Javier;Gutierrez, Santiago;Kosalkova, Katarina;Castillo, Nancy-Isabel;Martin, Juan-F.
    • Journal of Microbiology and Biotechnology
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    • v.11 no.5
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    • pp.812-818
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    • 2001
  • Alginate and alginate-derived oligomannuronate enhanced penicillin production in shake flask and fermentor cultures of Penicillium chrysogenum Wis 54-1255 (containing a single copy of the penicillin gene cluster) and in the high producter strain P. chrysogenum AS-P-99 (containing multiple copies of the penicillin gene cluster). Alginate was not used as a single carbon source by P. chryogenum. The stimulatory effect on penicillin production was observed in a defined medium and, to a lower extent, in a complex production medium containing corn steep liquor. Alginate-supplemented cells showed higher transcript levels of the three penicillin biosynthetic genes, pcbAB, pcbC, and penDE, than cells grown in the absence of alginate. The promoters of the pcbAB, pcbC, and penDE genes were coupled to the reporter lacZ gene and introduced as monocopy constructions in P. chrysogenum Wis 54-1225 npe10 by targeted integration in the pyrG locus; the reporter ${\beta}$-galactosidase activity expressed from the three promoters was stimulated by alginate added to the culture medium of the transformants. These results indicate that the stimulation of penicillin production by alginate was derived from an increase in the transcriptional activity of the penicillin biosynthesis genes. The induction by alginate of the transcription of the three penicillin biosynthetic genes is good example of the coordinated induction of secondary metabolism genes by elicitors of plant (or microbial) origin.

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Biocomputational Characterization and Evolutionary Analysis of Bubaline Dicer1 Enzyme

  • Singh, Jasdeep;Mukhopadhyay, Chandra Sekhar;Arora, Jaspreet Singh;Kaur, Simarjeet
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.6
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    • pp.876-887
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    • 2015
  • Dicer, an ribonuclease type III type endonuclease, is the key enzyme involved in biogenesis of microRNAs (miRNAs) and small interfering RNAs (siRNAs), and thus plays a critical role in RNA interference through post transcriptional regulation of gene expression. This enzyme has not been well studied in the Indian water buffalo, an important species known for disease resistance and high milk production. In this study, the primary coding sequence (5,778 bp) of bubaline dicer (GenBank: AB969677.1) was determined and the bubaline Dicer1 biocomputationally characterized to determine the phylogenetic signature among higher eukaryotes. The evolutionary tree revealed that all the transcript variants of Dicer1 belonging to a specific species were within the same node and the sequences belonging to primates, rodents and lagomorphs, avians and reptiles formed independent clusters. The bubaline dicer1 is closely related to that of cattle and other ruminants and significantly divergent from dicer of lower species such as tapeworm, sea urchin and fruit fly. Evolutionary divergence analysis conducted using MEGA6 software indicated that dicer has undergone purifying selection over the time. Seventeen divergent sequences, representing each of the families/taxa were selected to study the specific regions of positive vis-$\grave{a}$-vis negative selection using different models like single likelihood ancestor counting, fixed effects likelihood, and random effects likelihood of Datamonkey server. Comparative analysis of the domain structure revealed that Dicer1 is conserved across mammalian species while variation both in terms of length of Dicer enzyme and presence or absence of domain is evident in the lower organisms.

Cloning and Characterization of the HSP70 Gene, and Its Expression in Response to Diapauses and Thermal Stress in the Onion Maggot, Delia antiqua

  • Chen, Bin;Kayukawa, Takumi;Monteiro, Antonia;Ishikawa, Yukio
    • BMB Reports
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    • v.39 no.6
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    • pp.749-758
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    • 2006
  • The cytosolic members of the HSP70 family of proteins play key roles in the molecular chaperone machinery of the cell. In the study we cloned and sequenced the full-length cDNA of Delia antiqua HSP70 gene, which is 2461 bp long and encodes 643 a.a. with a calculated molecular mass of 70,787 Da. We investigated gene copies of cytosolic HSP70 members of 4 insect species with complete genome available, and found that they are quite variable with species. In order to characterize this protein we carried out an alignment and a phylogenetic analysis with 41 complete protein sequences from insects. The analysis divided the cytosolic members of the family into two classes, HSP70 and HSC70, distinguishable on the basis of 15 residues. HSP70 class members were slightly shorter in length and smaller in molecular mass relative to the HSC70 class members, and the conservative and functional regions in these sequences were documented. Mainly, we investigated the expression of Delia antiqua HSP70 gene, in response to diapauses and thermal stresses. Both summer and winter diapauses elevated HSP70 transcript levels. Cold-stress led to increased HSP70 expression levels in summer- and winter-diapausing pupae, but heat-stress elevated the levels only in the winter-diapausing pupae. In all cases, the expression levels, after being elevated, gradually decreased with time. HSP70 expression was low in non-diapausing pupae but was up-regulated following cold- and heat-stresses. Heat-stress gradually increased the mRNA level with time whereas cold-stress gradually decreased levels after an initial increase.

LebZIP2 induced by salt and drought stress and transient overexpression by Agrobacterium

  • Seong, Eun-Soo;Kwon, Soon -ung;Ghimire, Bimal Kumar;Yu, Chang-Yeon;Cho, Dong-Ha;Lim, Jung-Dae;Kim, Kyoung-Su;Heo, Kweon;Lim, Eun-Sang;Chung, Ill-Min;Kim, Myong-Jo;Lee, Youn-Su
    • BMB Reports
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    • v.41 no.10
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    • pp.693-698
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    • 2008
  • The full-length cDNA of LebZIP2 (Lycopersicon esculentum bZIP2) encodes a protein of 164 amino acids and contains a N-terminal basic-region leucine zipper domain. Analysis of the deduced tomato LebZIP2 amino acid sequence revealed that it shares 85% sequence identity with both tobacco bZIP and pepper CcbZIP. LebZIP2 mRNA is expressed at a high level exclusively in flowers. Presently, LebZIP2 was strongly increased also following NaCl and mannitol treatments. No significant LebZIP2 expression was evident following cold treatment. Transient LebZIP2 overexpression resulted in increased NbNOA1 and NbNR transcript levels in Nicotiana benthamiana leaves. Our results indicate that LebZIP2 might play roles as an abiotic stress-signaling pathway and as a transcriptional regulator of the NbNOA1 or NbNR genes.