• Title/Summary/Keyword: gene localization

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Molecular Analyses of the Metallothionein Gene Family in Rice (Oryza sativa L.)

  • Zhou, Gongke;Xu, Yufeng;Li, Ji;Yang, Lingyan;Liu, Jin-Yuan
    • BMB Reports
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    • v.39 no.5
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    • pp.595-606
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    • 2006
  • Metallothioneins are a group of low molecular mass and cysteine-rich metal-binding proteins, ubiquitously found in most living organisms. They play an important role in maintaining intracellular metal homeostasis, eliminating metal toxification and protecting against intracellular oxidative damages. Analysis of complete rice genome sequences revealed eleven genes encoding putative metallothionein (OsMT), indicating that OsMTs constitute a small gene family in rice. Expression profiling revealed that each member of the OsMT gene family differs not only in sequence but also in their tissue expression patterns, suggesting that these isoforms may have different functions they perform in specific tissues. On the basis of OsMT structural and phylogenetic analysis, the OsMT family was classified as two classes and class I was subdivided into four types. Additionally, in this paper we also present a complete overview of this family, describing the gene structure, genome localization, upstream regulatory element, and exon/intron organization of each member in order to provide valuable insight into this OsMT gene family.

Overexpression of the Downward Leaf Curling (DLC) Gene from Melon Changes Leaf Morphology by Controlling Cell Size and Shape in Arabidopsis Leaves

  • Kee, Jae-Jun;Jun, Sang Eun;Baek, Seung-A;Lee, Tae-Soo;Cho, Myung Rae;Hwang, Hyun-Sik;Lee, Suk-Chan;Kim, Jongkee;Kim, Gyung-Tae;Im, Kyung-Hoan
    • Molecules and Cells
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    • v.28 no.2
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    • pp.93-98
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    • 2009
  • A plant-specific gene was cloned from melon fruit. This gene was named downward leaf curling (CmDLC) based on the phenotype of transgenic Arabidopsis plants overexpressing the gene. This expression level of this gene was especially upregulated during melon fruit enlargement. Overexpression of CmDLC in Arabidopsis resulted in dwarfism and narrow, epinastically curled leaves. These phenotypes were found to be caused by a reduction in cell number and cell size on the adaxial and abaxial sides of the epidermis, with a greater reduction on the abaxial side of the leaves. These phenotypic characteristics, combined with the more wavy morphology of epidermal cells in overexpression lines, indicate that CmDLC overexpression affects cell elongation and cell morphology. To investigate intracellular protein localization, a CmDLC-GFP fusion protein was made and expressed in onion epidermal cells. This protein was observed to be preferentially localized close to the cell membrane. Thus, we report here a new plant-specific gene that is localized to the cell membrane and that controls leaf cell number, size and morphology.

Expression of peroxiredoxin I regulated by gonadotropins in the rat ovary

  • Lee, Yu-Il;Kang, Woo-Dae;Kim, Mi-Young;Cho, Moon-Kyoung;Chun, Sang-Young
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.1
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    • pp.18-23
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    • 2011
  • Objective: Peroxiredoxins (Prxs) play an important role in regulating cellular differentiation and proliferation in several types of mammalian cells. This report examined the expression of Prx isotype I in the rat ovary after hormone treatment. Methods: Immature rats were injected with 10 IU of pregnant mare's serum gonadotropin (PMSG) to induce the growth of multiple preovulatory follicles and 10 IU of human chorionic gonadotropin (hCG) to induce ovulation. Immature rats were also treated with diethylstilbestrol (DES), an estrogen analogue, to induce the growth of multiple immature follicles. Northern blot analysis was performed to detect gene expression. Cell-type specific localization of Prx I mRNA were detected by in situ hybridization analysis. Results: During follicle development, ovarian Prx I gene expression was detected in 3-day-old rats and had increased in 21-day-old rats. The levels of Prx I mRNA slightly declined one to two days following treatment with DES. A gradual increase in Prx I gene expression was observed in ovaries obtained from PMSG-treated immature rats. Furthermore, hCG treatment of PMSG-primed rats resulted in a gradual stimulation of Prx I mRNA levels by 24 hours (2.1-fold increase) following treatment, which remained high until 72 hours following treatment. In situ hybridization analysis revealed the expression of the Prx I gene in the granulosa cells of PMSG-primed ovaries and in the corpora lutea of ovaries stimulated with hCG for 72 hours. Conclusion: These results demonstrate the gonadotropin and granulosa cell-specific stimulation of Prx I gene expression, suggesting its role as a local regulator of follicle development.

Annexin A2 gene interacting with viral matrix protein to promote bovine ephemeral fever virus release

  • Chen, Lihui;Li, Xingyu;Wang, Hongmei;Hou, Peili;He, Hongbin
    • Journal of Veterinary Science
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    • v.21 no.2
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    • pp.33.1-33.15
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    • 2020
  • Bovine ephemeral fever virus (BEFV) causes bovine ephemeral fever, which can produce considerable economic damage to the cattle industry. However, there is limited experimental evidence regarding the underlying mechanisms of BEFV. Annexin A2 (AnxA2) is a calcium and lipid-conjugated protein that binds phospholipids and the cytoskeleton in a Ca2+-dependent manner, and it participates in various cellular functions, including vesicular trafficking, organization of membrane domains, and virus proliferation. The role of the AnxA2 gene during virus infection has not yet been reported. In this study, we observed that AnxA2 gene expression was up-regulated in BHK-21 cells infected with the virus. Additionally, overexpression of the AnxA2 gene promoted the release of mature virus particles, whereas BEFV replication was remarkably inhibited after reducing AnxA2 gene expression by using the small interfering RNA (siRNA). For viral proteins, overexpression of the Matrix (M) gene promotes the release of mature virus particles. Moreover, the AnxA2 protein interaction with the M protein of BEFV was confirmed by GST pull-down and co-immunoprecipitation assays. Experimental results indicate that the C-terminal domain (268-334 aa) of AxnA2 contributes to this interaction. An additional mechanistic study showed that AnxA2 protein interacts with M protein and mediates the localization of the M protein at the plasma membrane. Furthermore, the absence of the AnxA2-V domain could attenuate the effect of AnxA2 on BEFV replication. These findings can contribute to elucidating the regulation of BEFV replication and may have implications for antiviral strategy development.

Molecular Cloning and Characterization of a Heat Shock Protein 70-related cDNA from Olive flounder (Paralichthys olivaceus)

  • Kim, Woo-Jin;Lee, Jeong-Ho;Kim, Kyung-Kil;Lee, Sang-Jun;Kang, Ho-Sung;Kim, Han-Do
    • Journal of Aquaculture
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    • v.12 no.2
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    • pp.91-100
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    • 1999
  • The complete nucleotide sequence of olive flounder (Paralichthys olivaceus) hsp70-related rDNA was determined by RT- and RACE-PCR methods. A full-length of hsp70-related cDNA has an open reading frame of 1.95 kb encoding 650 amino acids with a calculated molecular weight of 71.1 kD. A corresponding hsp70-related protein contains a number of conserved elements including an ATP-binding domain, a nuclear localization signal and the carboxyl terminal motif, EEVD, which may have a role in chaperone function. Comparison of nucleotide and predicted amino acid sequence between olive flounder hsp70-related gene and hsp/hsc70 genes of other species revealed a high similarity with the cognate form of these genes. These results indicated that we recovered likely to be a olive flounder cognate hsc70 gene.

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Contribution of RIZ1 to Regulation of Proliferation and Migration of a Liver Fluke-Related Cholangiocarcinoma Cell

  • Khaenam, Prasong;Niibori, Akiko;Okada, Seiji;Jearanaikoon, Patcharee;Araki, Norie;Limpaiboon, Temduang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.4007-4011
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    • 2012
  • Purpose: Retinoblastoma-interacting zinc finger gene (RIZ1) is a tumor suppressor gene which is highly inactivated by promoter hypermethylation in patients with liver fluke-related cholangiocarcinoma (CCA). Epigenetic aberration of this gene might withdraw the ability to restrain tumor cell proliferation and migration. We aimed to define the role of RIZ1 on cell proliferation and migration in CCA cell line. Materials and methods: Small interference RNA (siRNA) was used to knock down the expression of RIZ1 in a CCA-derived cell line in which cell proliferation and cell migration were performed. Results: A predominant nuclear localization of RIZ1 was observed. Reduction of RIZ1 by siRNA augmented cell proliferation and migration. Conclusion: The result suggested that RIZ1 might play a role in regulating cell proliferation and migration in CCA. Reduction of RIZ1 expression may aggravate the progression of CCA.

Differential expression of soybean SLTI100 gene encoding translation elongation factor 1A by abiotic stresses

  • Chung, Eun-Sook;Cho, Chang-Woo;So, Hyun-A;Yun, Bo-Hyun;Lee, Jai-Heon
    • Journal of Plant Biotechnology
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    • v.36 no.3
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    • pp.255-260
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    • 2009
  • The translation elongation factor 1A, eEF1A, catalyzes the binding of aminoacyl-tRNA to the A-site of the ribosome by a GTP-dependent mechanism. By subtractive suppression hybridization technique, we have isolated a soybean low-temperature inducible gene, SLTI100 encoding translation elongation factor 1A. Multiple sequence alignments and phylogenic analysis showed that SLTI100 and other eEF1As originated from diverse organisms are highly conserved. RNA expression of SLTI100 was specifically induced by low temperature, high salt, ABA, or drought stress. Based on the subcellular localization of the corresponding gene product fused to GFP, we were able to confirm that SLTI100-GFP was restricted to the nucleus and cytoplasm. We propose that soybean eEF1A may play an important role in translational regulation during abiotic stress responses in plants.

m6A in the Signal Transduction Network

  • Jang, Ki-Hong;Heras, Chloe R.;Lee, Gina
    • Molecules and Cells
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    • v.45 no.7
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    • pp.435-443
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    • 2022
  • In response to environmental changes, signaling pathways rewire gene expression programs through transcription factors. Epigenetic modification of the transcribed RNA can be another layer of gene expression regulation. N6-adenosine methylation (m6A) is one of the most common modifications on mRNA. It is a reversible chemical mark catalyzed by the enzymes that deposit and remove methyl groups. m6A recruits effector proteins that determine the fate of mRNAs through changes in splicing, cellular localization, stability, and translation efficiency. Emerging evidence shows that key signal transduction pathways including TGFβ (transforming growth factor-β), ERK (extracellular signal-regulated kinase), and mTORC1 (mechanistic target of rapamycin complex 1) regulate downstream gene expression through m6A processing. Conversely, m6A can modulate the activity of signal transduction networks via m6A modification of signaling pathway genes or by acting as a ligand for receptors. In this review, we discuss the current understanding of the crosstalk between m6A and signaling pathways and its implication for biological systems.

BRCA1 Protein Was Not Expressed in a Normal Human Breast Epithelial Cell Type With Stem Cell and Luminal Characteristics

  • Kang, Kyung-Sun;Maki Saitoh;Angelar Cruz;Chan, Chia-Cheng;Cho, Jae-Jin
    • Toxicological Research
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    • v.14 no.2
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    • pp.123-127
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    • 1998
  • BRCA1 is a tumor suppresser gene in familial cases of breast cancer. It has been controversial whether the subcellular localization of BRCA1 is located in nuclei or cytoplasm in normal human breast cells. We found that a p220 protein was expressed in Type II Normal human breast epithelial cells (NHBEC) but not in Type I NHBEC in Western blot analysis using the 17F8 (3A2) antibody. Immunostaining using the same antibody revealed positive staining in nuclei, cytoplasm and perinuclei of Type II cells and negative staining in Type I NHBEC. The p220 protein, however, was expressed in SV40 immortalized Type I NHBEC and tumorigenic cells derived from them after x-ray and neu oncogene treatment. The subcelluar localization was mostly cytoplasmic and punctate in the nuclei. The breast carcinoma cell lines, MCF-7 and T47D, also expressed the p220 protein. Using RT-PCR, we observed the expression of BRCA1 mRNA in both Type I and Type II NHBEC. This result indicated that there might be mechanisms involved in post-translational or translational regulation of BRCA1 gene. It is speculated that the absence of BRCA1 protein expression in Type I NHBEC might playa role in their susceptibility to neoplastic transformation.

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Entomocidal Protein Gene Localization of Bacillus thuringiensis serovar. kurstaki HD73 and Isolates KBS722 (Bacillus thuringiensis serovar. kurstaki HD73균과 분리균 KBS722의 곤충치사 내독소 단백질의 Gene localization에 관한 연구)

  • 오상수;박영남;구본성;박유신;윤상홍
    • Microbiology and Biotechnology Letters
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    • v.17 no.2
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    • pp.142-147
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    • 1989
  • Six plasmids of B. thuringiensis serovar. kurstaki HD73 were detected, with approximate sizes of 7.4, 7.8, 8.1, 11.3, and 75 Kb, as well as a low copied plasmid of similar length to 75 Kb. Partially cured mutants from B. thuringiensis HD73 were obtained either by the treatment of the curing agent, ethidium bromide(0.02 $\mu\textrm{g}$/$m\ell$) or by spontaneous curing, Acrystalliferous mutants(Cry$^-$) were identified by microscopic observation and immunoblotting with polyclonal antibody against 133 KD deltaendotoxin of HD73. Ten Cry$^-$ mutants were found to be lack of 75 Kb plasmid. These results implicated that this plasmid was associated with delta-endotoxin production, After isolating the mutants, we streaked them on potato dextrose agar, spizizen casamino acid glucose, starch agar, and nutrient agar. Only on starch agar medium did morphologies of Cry$^-$ appear translucent and light greyish. On the other hand, the mutants of B. thuringiensis isolated from Korean soil, designated KBS722, were obtained by the treatment of novobiocin (3 $\mu\textrm{g}$/$m\ell$). Acrystalliferous mutants of KBS722 were less translucent than HD73 mutants' only on nutrient agar medium. Compared the plasmid profile of the mutants with delta-endotoxin production, the results seemed to indicate that the insecticidal protein gene of B. thuringiensis isolates KBS722 located on about 225 Kb plasmid DNA.

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