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FEBS Open Bio

Si-Jia Chen, Wen Wang, Feng-Yi Zhang, Yan-Long Jia, Xiao-Yin Wang, Xiao Guo, Shao-Nan Chen, Jian-Hui Gao, Tian-Yun Wang
Chinese hamster ovary (CHO) cells are one of the most commonly used expression systems for the production of recombinant proteins but low levels of transgene expression and transgene silencing are frequently encountered. Epigenetic regulatory elements such as the chicken β-globin locus control region hypersensitive site 4 (HS4) and scaffold/matrix attachment regions (S/MARs) have positive effects on transgene expression. In this study, a chimeric HS4-SAR was cloned upstream or downstream of an enhanced green fluorescent protein (eGFP) expression cassette in a eukaryotic vector, and the resulting vectors were transfected into CHO cells...
December 2017: FEBS Open Bio
Xingang Jia, Yisu Liu, Qiuhong Han, Zuhong Lu
Analysis of gene expression data by clustering and visualizing played a central role in obtaining biological knowledge. Here, we used Pearson's correlation coefficient of multiple-cumulative probabilities (PCC-MCP) of genes to define the similarity of gene expression behaviors. To answer the challenge of the high-dimensional MCPs, we used icc-cluster, a clustering algorithm that obtained solutions by iterating clustering centers, with PCC-MCP to group genes. We then used t-statistic stochastic neighbor embedding (t-SNE) of KC-data to generate optimal maps for clusters of MCP (t-SNE-MCP-O maps)...
December 2017: FEBS Open Bio
Takao Fuchigami, Hirofumi Koyama, Michiko Kishida, Yoshiaki Nishizawa, Mikio Iijima, Toshiro Kibe, Masahiro Ueda, Tohru Kiyono, Yoshimasa Maniwa, Norifumi Nakamura, Shosei Kishida
Ameloblastoma is a benign tumor of the odontogenic epithelium with several histological subtypes. All subtypes of ameloblastoma contain abundant stroma; the tumor cells invade collectively into the surrounding tissues without losing intratumor cell attachments. However, the molecular mechanisms mediating ameloblastoma invasion remain unclear. Here, we evaluated the functional significance of the interactions between ameloblastoma tumor cells and stromal fibroblasts on collective cellular invasion using a three-dimensional cultivation method, double-layered collagen gel hemisphere (DL-CGH) culture...
December 2017: FEBS Open Bio
Yuka Sone, Shimpei Uraguchi, Yasukazu Takanezawa, Ryosuke Nakamura, Hidemitsu Pan-Hou, Masako Kiyono
Bacterial resistance to mercury compounds (mercurials) is mediated by proteins encoded by mercury resistance (mer) operons. Six merE variants with site-directed mutations were constructed to investigate the roles of the cysteine and histidine residues in MerE protein during mercurial transport. By comparison of mercurial uptake by the cell with intact and/or variant MerE, we showed that the cysteine pair in the first transmembrane domain was critical for the transport of both Hg(II) and CH 3Hg(I). Also, the histidine residue located near to the cysteine pair was critical for Hg(II) transport, whereas the histidine residue located on the periplasmic side was critical for CH 3Hg(I) transport...
December 2017: FEBS Open Bio
Paweł Kozielewicz, Hatun Alomar, Syaratul Yusof, Gillian Grafton, Alison J Cooper, S John Curnow, James W Ironside, Hardev Pall, Nicholas M Barnes
A number of members of the G protein-coupled receptor class of cell surface receptors are 'orphans' with no known endogenous ligand. One of these orphan receptors is GPR61; there are little data about its expression in human cells and tissues. In this study, we investigated the post-translational modification of GPR61 by N-glycosylation at an identified consensus N-glycosylation site (N12) and the impact of this modification upon the subcellular expression of the protein. The N-glycosylation inhibitor tunicamycin reduced the apparent molecular weight of immunoreactivity associated with myc-tagged GPR61 by 1-2 kDa, which was comparable to the evident molecular weight of the myc-tagged N12S GPR61 mutant with disrupted consensus N-glycosylation site...
December 2017: FEBS Open Bio
Tsukasa Tominari, Ryota Ichimaru, Shosei Yoshinouchi, Chiho Matsumoto, Kenta Watanabe, Michiko Hirata, Florian M W Grundler, Masaki Inada, Chisato Miyaura
(-)-Epigallocatechin-3-O-gallate (EGCG), present in green tea, exhibits antioxidant and antiallergy effects. EGCG3″Me, a 3-O-methylated derivative of EGCG, has been reported to show similar biological functions; the inhibitory activity of EGCG3″Me in a mouse allergy model was more potent than that of EGCG, probably due to the efficiency of absorption from the intestine. However, the functional potency of these EGCGs is controversial in each disease model. We previously observed that EGCG suppressed inflammatory bone resorption and prevented alveolar bone loss in a mouse model of periodontosis...
December 2017: FEBS Open Bio
Chuan-Fu Tsai, Hsin-Yi Lin, Wei-Li Hsu, Ching-Hsiu Tsai
The nonstructural protein 1 (NS1) of the influenza A virus (IAV) is a multifunctional protein that counteracts host cell antiviral responses and inhibits host cell pre-mRNA processing. NS1 contains two nuclear localization signals that facilitate NS1 shuttling between cytoplasm and nucleus. In this study, we initially observed the novel mitochondria localization of NS1 in a subset of transfected cells. We then further monitored the localization dynamics of the NS1 protein in live cells infected with IAV expressing NS1 with insertion of a tetracysteine-tag...
December 2017: FEBS Open Bio
Jin-Lei Sui, Xiao-Hu Xiao, Ji-Yan Qi, Yong-Jun Fang, Chao-Rong Tang
SWEET proteins play an indispensable role as a sugar efflux transporter in plant development and stress responses. The SWEET genes have previously been characterized in several plants. Here, we present a comprehensive analysis of this gene family in the rubber tree, Hevea brasiliensis. There are 36 members of the SWEET gene family in this species, making it one of the largest families in plant genomes sequenced so far. Structure and phylogeny analyses of these genes in Hevea and in other species demonstrated broad evolutionary conservation...
December 2017: FEBS Open Bio
Kazumi Hirano, Masakazu Namihira
Flavin adenine dinucleotide (FAD), synthesized from riboflavin, is redox cofactor in energy production and plays an important role in cell survival. More recently, riboflavin deficiency has been linked to developmental disorders, but its role in stem cell differentiation remains unclear. Here, we show that FAD treatment, using DMSO as a solvent, enabled an increase in the amount of intracellular FAD and promoted neuronal differentiation of human neural stem cells (NSCs) derived not only from fetal brain, but also from induced pluripotent stem cells...
December 2017: FEBS Open Bio
Shujun Wang, Yaling Dou, Hong Yang, Anping Ni, Rui Zhang, Jiaming Qian
Cytomegalovirus (CMV) infection is associated with glucocorticoid resistance in ulcerative colitis (UC) and may exacerbate the disease course. However, the underlying pathogenicity remains unclear. The aim of this study was to explore possible underlying mechanisms during CMV latency and lytic infection in the human mononuclear cell line THP-1. Latent and activated CMV infection cell models were established. We performed real-time PCR and western blotting to examine changes in glucocorticoid receptors (GRs) during CMV latency and activation...
December 2017: FEBS Open Bio
Hannah Berberich, Felix Terwesten, Sinja Rakow, Peeyush Sahu, Caroline Bouchard, Marion Meixner, Sjaak Philipsen, Peter Kolb, Uta-Maria Bauer
Protein arginine methyltransferase 4 (PRMT4) is an essential epigenetic regulator of fundamental and conserved processes during vertebrate development, such as pluripotency and differentiation. Surprisingly, PRMT4 homologs have been identified in nearly all vertebrate classes except the avian genome. This raises the possibility that in birds PRMT4 functions are taken over by other PRMT family members. Here, we reveal the existence of a bona fide PRMT4 homolog in the chicken, Gallus gallus. Using a biochemical approach, we initially purified a putative chicken PRMT4 protein and thus provided the first evidence for the presence of an endogenous PRMT4-specific enzymatic activity toward histone H3 arginine 17 (H3R17) in avian cells...
December 2017: FEBS Open Bio
Xiaotong Sun, Jing Wei, Yawei Tang, Bing Wang, Yan Zhang, Lei Shi, Jianping Guo, Fanlei Hu, Xia Li
Rheumatoid arthritis (RA) is a progressive autoimmune disease affecting the joints. In this study, we investigated the role of the pro-angiogenic factor leptin in regulating reactive oxygen species (ROS) to promote cell migration and angiogenesis in RA. We showed that leptin triggered RA fibroblast-like synoviocyte (FLS) migration by increased ROS expression. Additionally, leptin enhanced human umbilical vein endothelial cell (HUVEC) tube formation in a ROS/hypoxia-inducible factor-1α-dependent manner, accompanied by increased production of vascular endothelial growth factor and interleukin (IL)-6...
December 2017: FEBS Open Bio
Rattawan Poodproh, Supavadee Kaewmeechai, Ladda Leelawatwattana, Porntip Prapunpoj
Transthyretin (TTR) is a transporter for thyroid hormone (TH) and retinol, the latter via binding with retinol binding protein (RBP). Both the N-terminal and C-terminal regions of the TTR subunit are located in close proximity to the central binding channel for ligands. During the evolution of vertebrates, these regions changed in length and hydropathy. The changes in the N-terminal sequence were demonstrated to affect the binding affinities for THs and RBP. Here, the effects of changes in the C-terminal sequence were determined...
December 2017: FEBS Open Bio
Fangwei Li, Wenhua Shi, Yixin Wan, Qingting Wang, Wei Feng, Xin Yan, Jian Wang, Limin Chai, Qianqian Zhang, Manxiang Li
The expression of microRNA (miR)-140-5p is known to be reduced in both pulmonary arterial hypertension (PAH) patients and monocrotaline-induced PAH models in rat. Identification of target genes for miR-140-5p with bioinformatics analysis may reveal new pathways and connections in PAH. This study aimed to explore downstream target genes and relevant signaling pathways regulated by miR-140-5p to provide theoretical evidences for further researches on role of miR-140-5p in PAH. Multiple downstream target genes and upstream transcription factors (TFs) of miR-140-5p were predicted in the analysis...
December 2017: FEBS Open Bio
Natsaran Saichana, Katsuyuki Tanizawa, Hiroshi Ueno, Jiří Pechoušek, Petr Novák, Jitka Frébortová
PqqE is a radical S-adenosyl-l-methionine (SAM) enzyme that catalyzes the initial reaction of pyrroloquinoline quinone (PQQ) biosynthesis. PqqE belongs to the SPASM (subtilosin/PQQ/anaerobic sulfatase/mycofactocin maturating enzymes) subfamily of the radical SAM superfamily and contains multiple Fe-S clusters. To characterize the Fe-S clusters in PqqE from Methylobacterium extorquens AM1, Cys residues conserved in the N-terminal signature motif (CX 3 CX 2C) and the C-terminal seven-cysteine motif (CX 9-15 GX 4 CX n CX 2 CX 5 CX 3 CX n C; n = an unspecified number) were individually or simultaneously mutated into Ser...
December 2017: FEBS Open Bio
Manabu Koike, Yasutomo Yutoku, Aki Koike
Molecularly targeted therapies have high specificity and significant cancer-killing effect. However, their antitumor effect might be greatly diminished by variation in even a single amino acid in the target site, as it occurs, for example, as a consequence of SNPs. Increasing evidence suggests that the DNA repair protein Ku80 is an attractive target molecule for the development of next-generation radiosensitizers for human cancers. However, the localization, post-translational modifications (PTMs), and complex formation of Ku80 have not been elucidated in canines...
December 2017: FEBS Open Bio
Marta Viano, Daniela Alotto, Alessia Aillon, Carlotta Castagnoli, Francesca Silvagno
During their spatial and differentiative progression, keratinocytes face a thermal gradient, from 37 °C in the proliferating basal layer to 32 °C found in skin surface. In our study, we hypothesized that this difference in temperature must be balanced by increasing the heat produced during respiratory activity. We demonstrated that at 33 °C human primary keratinocytes and HaCaT cells raised mitochondrial energy metabolism, but not glycolytic activity. At 33 °C, the increased mitochondrial ATP synthesis was associated with a strong induction of the modulator of the respiratory chain estrogen receptor β, whereas uncoupling protein 1 expression was not changed...
December 2017: FEBS Open Bio
Prasad Kasaragod, Getnet B Midekessa, Shruthi Sridhar, Werner Schmitz, Tiila-Riikka Kiema, Jukka K Hiltunen, Rik K Wierenga
Multifunctional enzyme, type-1 (MFE1) is a monomeric enzyme with a 2E-enoyl-CoA hydratase and a 3S-hydroxyacyl-CoA dehydrogenase (HAD) active site. Enzyme kinetic data of rat peroxisomal MFE1 show that the catalytic efficiencies for converting the short-chain substrate 2E-butenoyl-CoA into acetoacetyl-CoA are much lower when compared with those of the homologous monofunctional enzymes. The mode of binding of acetoacetyl-CoA (to the hydratase active site) and the very similar mode of binding of NAD + and NADH (to the HAD part) are described and compared with those of their monofunctional counterparts...
December 2017: FEBS Open Bio
(no author information available yet)
[This corrects the article DOI: 10.1002/2211-5463.12286.].
November 2017: FEBS Open Bio
Shimon Takagi, Fumitaka Momose, Yuko Morikawa
The Gag protein of HIV multimerizes to form viral particles. The GagPol protein encoding virus-specific enzymes, such as protease, reverse transcriptase, and integrase, is incorporated into HIV particles via interactions with Gag. The catalytically active forms of these enzymes are dimeric or tetrameric. We employed Förster resonance energy transfer (FRET) assays to evaluate Gag-Gag, Gag-GagPol, and GagPol-GagPol interactions and investigated Gag and Pol interdomains tolerant to fluorescent protein insertion for FRET assays...
November 2017: FEBS Open Bio
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