• Title/Summary/Keyword: ortholog

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Identification and Functional Analysis of SEDL-binding and Homologue Proteins by Immobilized GST Fusion and Motif Based Methods

  • Hong, Ji-Man;Jeong, Mi-Suk;Kim, Jae-Ho;Kim, Boog-il;Holbrook, Stephen R.;Jang, Se-Bok
    • Bulletin of the Korean Chemical Society
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    • v.29 no.2
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    • pp.381-388
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    • 2008
  • An X-linked skeletal disorder, SEDT (spondyloepiphyseal dysplasia tarda) is a genetic disease characterized by a disproportionately short trunk and short stature caused by mutations in the SEDL gene. This gene is evolutionarily conserved from yeast to human. The yeast SEDL protein ortholog, Trs20p, has been isolated as a member of a large multi-protein complex called the transport protein particle (TRAPP), which is involved in endoplasmic reticulum (ER)-to-Golgi transport. The interaction between SEDL and partner proteins is important in order to understand the molecular mechanism of SEDL functions. We isolated several SEDL-binding proteins derived from rat cells by an immobilized GST-fusion method. Furthermore, the SEDL-homologue proteins were identified using motif based methods. Common motifs between SEDL-binding proteins and SEDL-homologue proteins were classified into seven types and 78 common motifs were revealed. Sequence similarities were contracted to seven types using phylogenetic trees. In general, types I-III and VI were classified as having the function of acetyl-CoA carboxylase, glycogen phosphorylase, isocitrate dehydrogenase, and enolase, respectively, and type IV was found to be functionally related to the GST protein. Types V and VII were found to contribute to TRAPP vesicle trafficking.

Cloning and Nucleotide Sequence Analysis of the rpoH Gene from Methylovorus sp. Strain SS1 DSM11726 (Methylovorus sp. Strain SS1 DSM11726으로부터 rpoH 유전자의 클로닝과 염기서열 분석)

  • Eom, Chi-Yong;Song, Seung-Eun;Park, Mi-Hwa;Kim, Young-Min
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.177-183
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    • 2007
  • Using complementation of RpoH deficient E. coli strain A7448, the rpoH gene encoding heat shock sigma factor 32 (${\sigma}^{32}$) from Methylovorus sp. strain SS1 DSM11726 was cloned and sequenced. Sequence analysis of a stretch of 1,796-bp revealed existence of an open reading frame encoding a polypeptide of 284 amino acid (32,006 dalton). Deduced amino acid sequence of the Methylovorus sp. strain SS1 RpoH showed that 59.6%, 39.1% and 51.4% identities with those of Nitrosomonas europaea (${\beta}$-proteobacteria), Agrobacterium tumefaciens ($\alpha$-proteobacteria) and E. coli (${\gamma}$-proteobacteria). The expression level of the functional ortholog of RpoH of Methylovorus sp. strain SS1 was increased transiently after heat induction, further indicating that it functions as a heat shock sigma factor.

Negative Role of wblA in Response to Oxidative Stress in Streptomyces coelicolor

  • Kim, Jin-Su;Lee, Han-Na;Kim, Pil;Lee, Heung-Shick;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.736-741
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    • 2012
  • In this study, we analyzed the oxidative stress response of wblA ($\underline{w}$hi$\underline{B}$-$\underline{l}$ike gene $\underline{A}$, SCO3579), which was previously shown to be a global antibiotic down-regulator in Streptomyces coelicolor. Ever since a WblA ortholog named WhcA in Corynebacterium glutamicum was found to play a negative role in the oxidative stress response, S. coelicolor wblA has been proposed to have a similar effect. A wblA-deletion mutant exhibited a less sensitive response to oxidative stress induced by diamide present in solid plate culture. Using real-time RT-PCR analysis, we also compared the transcription levels of oxidative stress-related genes, including sodF, sodF2, sodN, trxB, and trxB2, between S. coelicolor wild type and a wblA-deletion mutant in the presence or absence of oxidative stress. Target genes were expressed higher in the wblA-deletion mutant compared with wild type, both in the absence and presence of oxidative stress. Moreover, expression of these target genes in S. coelicolor wild type was stimulated only in the presence of oxidative stress, suggesting that WblA plays a negative role in the oxidative stress response of S. coelicolor, similar to that of C. glutamicum WhcA, through the transcriptional regulation of oxidative stress-related genes.

Galactinol is Involved in Induced Systemic Resistance against Bacterial Infection and Environmental Stresses

  • Cho, Song-Mi;Kim, Su-Hyun;Kim, Young-Cheol;Yang, Kwang-Yeol;Kim, Kwang-Sang;Choi, Yong-Soo;Cho, Baik-Ho
    • Korean Journal of Plant Resources
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    • v.23 no.3
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    • pp.248-255
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    • 2010
  • We previously demonstrated that root colonization of the rhizobacterium, Pseudomonas chlororaphis O6, induced expression of a galactinol synthase gene (CsGolS1), and resulting galactinol conferred induced systemic resistance (ISR) against fungal and bacterial pathogens in cucumber leaves. To examine the role of galactinol on ISR, drought or high salt stress, we obtained T-DNA insertion Arabidopsis mutants at the AtGolS1 gene, an ortholog of the CsGolS1 gene. The T-DNA insertion mutant compromised resistance induced by the O6 colonization against Erwinia carotovora. Pharmaceutical application of 0.5 - 5 mM galactinol on roots was sufficient to elicit ISR in wild-type Arabidopsis against infection with E. carotovora. The involvement of jasmonic acid (JA) signaling on the ISR was validated to detect increased expression of the indicator gene PDF1.2. The T-DNA insertion mutant also compromised tolerance by increasing galactinol content in the O6-colonized plant against drought or high salt stresses. Taken together, our results indicate that primed expression of the galactinol synthase gene AtGolS1in the O6-colonized plants can play a critical role in the ISR against infection with E. carotovora, and in the tolerance to drought or high salt stresses.

Effects of Sus1, a component of TREX-2 complex, on growth and mRNA export in fission yeast (분열효모에서 TREX-2 복합체의 구성요소인 Sus1이 생장 및 mRNA 방출에 미치는 영향)

  • Bae, Soo Jeong;Yoon, Jin Ho
    • Korean Journal of Microbiology
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    • v.53 no.1
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    • pp.49-54
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    • 2017
  • Sus1 / ENY2 is a tiny conserved protein that is involved in chromatin remodeling and mRNA biogenesis. Sus1 is associated to two nuclear complexes, the transcriptional coactivator SAGA and the nuclear pore associated TREX2. In fission yeast, Schizosaccharomyces pombe, ortholog of Sus1 / ENY2 was identified from the genome database. Tetrad analysis showed that the S. pombe sus1 is not essential for growth. However, deletion of the sus1 gene caused cold-sensitive growth retardation with slight accumulation of $poly(A)^+$ RNA in the nucleus. And the Sus1-GFP protein is localized mainly in the nucleus. Yeast two-hybrid analysis and co-immunoprecipitation experiment showed that Sus1 interacts with Sac3, another subunit of TREX2 complex. These results suggest that S. pombe Sus1 is also involved in mRNA export from the nucleus as a component of TREX-2 complex.

Homologous Expression and T3SS-Dependent Secretion of TAP-Tagged Xo2276 in Xanthomonas oryzae pv. oryzae Induced by Rice Leaf Extract and Its Direct In Vitro Recognition of Putative Target DNA Sequence

  • Kim, Seunghwan;Nguyen, Thi-Dieu-Hanh;Lee, Joohee;Hong, Myoung-Ki;Pham, Tan-Viet;Ahn, Yeh-Jin;Lee, Byoung-Moo;Han, Ye Sun;Kim, Dong-Eun;Kim, Jeong-Gu;Kang, Lin-Woo
    • Journal of Microbiology and Biotechnology
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    • v.23 no.1
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    • pp.22-28
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    • 2013
  • Xo2276 is a putative transcription activator-like effector (TALE) in Xanthomonas oryzae pv. oryzae (Xoo). Xo2276 was expressed with a TAP-tag at the C-terminus in Xoo cells to enable quantitative analysis of protein expression and secretion. Nearly all TAP-tagged Xo2276 existed in an insoluble form; addition of rice leaf extracts from a Xoosusceptible rice cultivar, Milyang23, significantly stimulated secretion of TAP-tagged Xo2276 into the medium. In a T3SS-defective Xoo mutant strain, secretion of TAPtagged Xo2276 was blocked. Xo2276 is a Xoo ortholog of Xanthomonas campestris pv. vesicatoria (Xcv) AvrBs3 and contains a conserved DNA-binding domain (DBD), which includes 19.5 tandem repeats of 34 amino acids. Xo2276- DBD was expressed in E. coli and purified. Direct in vitro recognition of Xo2276-DBD on a putative target DNA sequence was confirmed using an electrophoretic mobility shift assay. This is the first study measuring the homologous expression and secretion of Xo2276 in vitro using rice leaf extract and its direct in vitro binding to the specific target DNA sequence.

The Rice FON1 Gene Controls Vegetative and Reproductive Development by Regulating Shoot Apical Meristem Size

  • Moon, Sunok;Jung, Ki-Hong;Lee, Do-Eun;Lee, Dong-Yeon;Lee, Jinwon;An, Kyungsook;Kang, Hong-Gyu;An, Gynheung
    • Molecules and Cells
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    • v.21 no.1
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    • pp.147-152
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    • 2006
  • Most plant organs develop from meristems. Rice FON1, which is an ortholog of Clv1, regulates stem cell proliferation and organ initiation. The point mutations, fon1-1 and fon1-2, disrupt meristem balance, resulting in alteration of floral organ numbers and the architecture of primary rachis branches. In this study, we identified two knockout alleles, fon1-3 and fon1-4, generated by T-DNA and Tos17 insertion, respectively. Unlike the previously isolated point mutants, the null mutants have alterations not only of the reproductive organs but also of vegetative tissues, producing fewer tillers and secondary rachis branches. The mutant plants are semi-dwarfs due to delayed leaf emergence, and leaf senescence is delayed. SEM analysis showed that the shoot apical meristems of fon1-3 mutants are enlarged. These results indicate that FON1 controls vegetative as well as reproductive development by regulating meristem size.

Nucleolar GTPase NOG-1 Regulates Development, Fat Storage, and Longevity through Insulin/IGF Signaling in C. elegans

  • Kim, Young-Il;Bandyopadhyay, Jaya;Cho, Injeong;Lee, Juyeon;Park, Dae Ho;Cho, Jeong Hoon
    • Molecules and Cells
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    • v.37 no.1
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    • pp.51-57
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    • 2014
  • NOG1 is a nucleolar GTPase that is critical for 60S ribosome biogenesis. Recently, NOG1 was identified as one of the downstream regulators of target of rapamycin (TOR) in yeast. It is reported that TOR is involved in regulating lifespan and fat storage in Caenorhabditis elegans. Here, we show that the nog1 ortholog (T07A9.9: nog-1) in C. elegans regulates growth, development, lifespan, and fat metabolism. A green fluorescence protein (GFP) promoter assay revealed ubiquitous expression of C. elegans nog-1 from the early embryonic to the adult stage. Furthermore, the GFP-tagged NOG-1 protein is localized to the nucleus, whereas the aberrant NOG-1 protein is concentrated in the nucleolus. Functional studies of NOG-1 in C. elegans further revealed that nog-1 knockdown resulted in smaller broodsize, slower growth, increased life span, and more fat storage. Moreover, nog-1 over-expression resulted in decreased life span. Taken together, our data suggest that nog-1 in C. elegans may be an important player in regulating life span and fat storage via the insulin/IGF pathway.

A Role for buttonhead in the Early Head and Trunk Development in the Beetle Tribolium castaneum

  • Jeon, Haewon;O, Jiyun;Jin, Sil;Lim, Jinsung;Choe, Chong Pyo
    • Development and Reproduction
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    • v.23 no.1
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    • pp.63-72
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    • 2019
  • The head gap gene buttonhead (btd) is required for the patterning of head segments in the early Drosophila embryo. Mutant phenotypes of btd display a gap-like phenotype in which antennal, intercalary, mandibular and the anterior portion of the maxillary segments are eliminated. In agreement with the phenotypes, btd is expressed in a stripe covering the head segments at the blastoderm stage. During the early phase of the germband extension, btd is expressed in stripes with single segmental periodicity, which is required for the formation of the peripheral nervous system. In contrast to the key role of btd in Drosophila embryonic development, it has been suggested that Tribolium ortholog of btd (Tc-btd) is dispensable for embryonic head development. In order for better understanding of the requirement of Tc-btd in the early Tribolium embryo, we re-analyzed the expression patterns and functions of Tc-btd during embryonic segmentation. Tc-btd is expressed in segmental stripes at the stages of blastoderm and germband elongation. Up to 28.3% of embryos in which Tc-btd is knocked down displays the loss of antennal, mandibular and the pregnathal regions in the head, with abdominal segments being disrupted in the trunk. Our findings suggest that Tc-btd is required for the head and trunk development in the early Tribolium embryo.

Blood transcriptome resources of chinstrap (Pygoscelis antarcticus) and gentoo (Pygoscelis papua) penguins from the South Shetland Islands, Antarctica

  • Kim, Bo-Mi;Jeong, Jihye;Jo, Euna;Ahn, Do-Hwan;Kim, Jeong-Hoon;Rhee, Jae-Sung;Park, Hyun
    • Genomics & Informatics
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    • v.17 no.1
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    • pp.5.1-5.9
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    • 2019
  • The chinstrap (Pygoscelis antarcticus) and gentoo (P. papua) penguins are distributed throughout Antarctica and the sub-Antarctic islands. In this study, high-quality de novo assemblies of blood transcriptomes from these penguins were generated using the Illumina MiSeq platform. A total of 22.2 and 21.8 raw reads were obtained from chinstrap and gentoo penguins, respectively. These reads were assembled using the Oases assembly platform and resulted in 26,036 and 21,854 contigs with N50 values of 929 and 933 base pairs, respectively. Functional gene annotations through pathway analyses of the Gene Ontology, EuKaryotic Orthologous Groups, and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases were performed for each blood transcriptome, resulting in a similar compositional order between the two transcriptomes. Ortholog comparisons with previously published transcriptomes from the $Ad{\acute{e}}lie$ (P. adeliae) and emperor (Aptenodytes forsteri) penguins revealed that a high proportion of the four penguins' transcriptomes had significant sequence homology. Because blood and tissues of penguins have been used to monitor pollution in Antarctica, immune parameters in blood could be important indicators for understanding the health status of penguins and other Antarctic animals. In the blood transcriptomes, KEGG analyses detected many essential genes involved in the major innate immunity pathways, which are key metabolic pathways for maintaining homeostasis against exogenous infections or toxins. Blood transcriptome studies such as this may be useful for checking the immune and health status of penguins without sacrifice.