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Genes to Cells: Devoted to Molecular & Cellular Mechanisms

Masashi Takao, Yutaka Nagai, Masami Ito, Tetsuhiko Ohba
Extracellular vesicles (EV) have attracted attention as circulating biomarkers for many diseases, particularly cancer. Conventional immunofluorescence staining has been used for detection of target antigens on EV by flow cytometry. However, the staining intensity depends on the amount of antigen expressed on the vesicles and is often only around the noise level. Instead of immunofluorescence, we combined immunomagnetic separation using nanosize MACS® MicroBeads with phospholipid staining of EV (IMS-PS method)...
September 16, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Yan Li, Kyosuke Kobayashi, Kosho Murayama, Kohichi Kawahara, Yuichi Shima, Akira Suzuki, Kenzaburo Tani, Atsushi Takahashi
FEAT, the protein encoded by methyltransferase like 13 (METTL13), is aberrantly upregulated in most human cancers and potently drives tumorigenesis in vivo; however, its role in normal tissues remains elusive. Immunoblotting has displayed weak FEAT expression in normal human tissues, including the testis. Here, we found that FEAT is expressed in fetal and adult Leydig cells in the testis. FEAT knockdown using siRNA increased primary cilia formation in MA-10 Leydig tumor cells, accompanied by enhanced 5' adenosine monophosphate-activated protein kinase (AMPK) activation...
September 4, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Mikio Takagi, Masaya Ikegawa, Takashi Shimada, Susumu Ishikawa, Mihoko Kajita, Takeshi Maruyama, Tomoko Kamasaki, Yasuyuki Fujita
At the initial stage of carcinogenesis, transformation occurs in single cells within the epithelium. Recent studies have revealed that the newly emerging transformed cells are often apically eliminated from epithelial tissues. However, the underlying molecular mechanisms of this cancer preventive phenomenon still remain elusive. In this study, we first demonstrate that myosin-II accumulates in Src-transformed cells when they are surrounded by normal epithelial cells. Knockdown of the heavy chains of myosin-II substantially diminishes apical extrusion of Src cells, suggesting that accumulated myosin-II positively regulates the apical elimination of transformed cells...
September 2, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Lijuan Huang, Masaaki Ozawa, Etsuko Miyamoto-Sato
To clarify the pathogenic mechanism of disease and establish effective therapies, animal disease models that can be dynamically analyzed are urgently required. Knockout mouse models and conditional genetically-engineered mouse models were developed to analyze genes and proteins involved in disease. However, these methods have drawbacks, including embryonic lethality, side effects, and low efficiency. To address this issue, we created a novel transgenic mouse model in which the YB1 gene was fused with a destabilizing domain (DD), named the YB1-DD mouse...
August 30, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Atsuo Iida, Zi Wang, Hiromi Hirata, Atsuko Sehara-Fujisawa
Integrins are transmembrane molecules that facilitate cell-to-cell and cell-to-extra cellular matrix (ECM) interactions. Integrin molecules are heterodimers that consist of an α- and β-subunits. The integrin β1 gene is widely expressed in vivo and is the major β molecule in many tissues; however, tissue specific roles of integrin β1 are still elusive. In this study, we investigated integrin β1 function in endothelial cells of zebrafish. An integrin β1b mutant zebrafish exhibited morphological abnormalities in blood vessel formation, cephalic hemorrhage, and a decreased responsiveness to tactile stimulation during development...
August 27, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Takahiro Komori, Atsushi Shibai, Hiromi Saito, Yuya Akeno, Arnaud Germond, Takaaki Horinouchi, Chikara Furusawa, Saburo Tsuru
Evolutionary strategies in growth improvement can be classified into r- or K-strategies. The former strategy corresponds to an evolutionary increase in growth rate, while the latter corresponds to an increase in the maximum amount of organisms or carrying capacity. What determines the strategies to be adopted during evolution? Spatial structures that compartmentalize the population into small patches are key to inducing the K-strategy. Interestingly, previous evolution experiments using Escherichia coli in a glucose-limited batch culture revealed that carrying capacity could improve evolutionally even in the absence of spatial structures...
August 24, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Yuki Hattori, Takaki Miyata
Neocortical development proceeds through the formation of new zones in which neural-lineage cells are organized based on their differentiation status. Although microglia initially distribute homogeneously throughout the growing cerebral wall, they accumulate in the inner cytogenic zone, the ventricular zone (VZ) and the subventricular zone (SVZ) in the mid-embryonic stage. However, the roles of these cells remain to be elucidated. In this study, we found that microglia, despite being only a minor population of the cells that constitute the cerebral wall, promote the differentiation of neural progenitor cells by frequently moving throughout the cortex; their migration is mediated by the CXCL12/CXCR4 system...
August 24, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Akihiro Nakamura, Natsuko Kawano, Kei Motomura, Akio Kuroda, Kiyoshi Sekiguchi, Mami Miyado, Kang Woojin, Yoshitaka Miyamoto, Maito Hanai, Maki Iwai, Mitsutoshi Yamada, Toshio Hamatani, Takakazu Saito, Hidekazu Saito, Mamoru Tanaka, Akihiro Umezawa, Kenji Miyado
In bacteria, a polymer of inorganic phosphate (Pi) (inorganic polyphosphate; polyP) is enzymatically produced and consumed as an alternative phosphate donor for adenosine triphosphate (ATP) production to protect against nutrient starvation. In vertebrates, polyP has been dismissed as a "molecular fossil" due to the lack of any known physiological function. Here, we have explored its possible role by producing transgenic mice (TG) widely expressing Saccharomyces cerevisiae exoplyphosphatase 1 (ScPPX1), which catalyzes hydrolytic polyP degradation...
August 24, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Rolf Müller, Maria Stumpf, Regina Wehrstedt, Salil K Sukumaran, Malte A Karow, Marija Marko, Angelika A Noegel, Ludwig Eichinger
phr2AB is the regulatory subunit of the Dictyostelium discoideum phosphatase PP2A and is the ortholog of the human B55 regulatory subunit of PP2A. phr2AB was isolated as a binding partner of the centrosomal protein CEP161, an ortholog of mammalian CDK5RAP2. CEP161 is presumably a phosphoprotein and a component of the Hippo pathway. The interaction site was located in the N-terminal half of CEP161 which encompasses the γTURC binding domain in CEP161. This binding domain is responsible for binding of the γ-tubulin ring complex which allows microtubule nucleation at the centrosome...
August 22, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Takehiko Yoko-O, Mariko Umemura, Akiko Komatsuzaki, Kazutaka Ikeda, Daisuke Ichikawa, Kumiko Takase, Noriyuki Kanzawa, Kazunobu Saito, Taroh Kinoshita, Ryo Taguchi, Yoshifumi Jigami
Yeasts have two classes of glycosylphosphatidylinositol (GPI) -anchored proteins; one is transferred to the cell wall while the other is retained on the plasma membrane. The lipid moieties of the GPI in Saccharomyces cerevisiae consist of either phosphatidylinositol (PI) or inositolphosphorylceramide (IPC). Cwh43p is involved in the remodeling of lipid from PI to IPC. We found that the GPI lipid moiety of Cwp2p in wild-type cells is PI. To elucidate the physiological role of the lipid remodeling by Cwh43p, we investigated the distribution of Gas1p and Cwp2p by immunoblotting, and found that Gas1p with the PI-form GPI lipid moiety in cwh43Δ mutant cells tends to be localized to the cell wall, suggesting that the IPC species in the GPI lipid moiety contributes to the retention of GPI-anchored proteins on the plasma membrane...
August 22, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Wei Wu, Kazutaka Sahara, Shoshiro Hirayama, Xian Zhao, Ayaka Watanabe, Jun Hamazaki, Hideki Yashiroda, Shigeo Murata
The proteasome core particle (CP) is a cytoplasmic and nuclear protease complex and is comprised of two α-rings and two β-rings stacked in order of αββα. The assembly of CP proceeds by ordered recruitment of β-subunits on an α-ring with help of assembly chaperones PAC1-PAC2, PAC3-PAC4, and UMP1. However, the mechanism of α-ring formation remains unsolved. Here, we show that α4, α5, α6, and α7 form a core intermediate as the initial process of α-ring assembly, which requires PAC3-PAC4. α1 and α3 can be incorporated independently into the core α4-α7 intermediate, whereas α2 incorporation is dependent on preceding incorporation of α1...
August 22, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Katsutoshi Maeda, Mitsuhiro Yoneda, Takeya Nakagawa, Kazuhiro Ikeda, Miki Higashi, Kaori Nakagawa, Mana Miyakoda, Katsuyuki Yui, Hiroaki Oda, Satoshi Inoue, Takashi Ito
Histone H2A phosphorylation plays a role both in chromatin condensation during mitosis and in transcriptional activation during the G1/S transition. Bub1 and NHK1/VRK1 have been identified as histone H2A kinases. However, little is known about the importance of histone H2A phosphorylation in chromosome segregation. Here, we expressed recombinant hBUB1 and confirmed that it phosphorylates histone H2A T120 in the in vitro-assembled nucleosome. Knockdown (KD) of BUB1 decreases bulk H2A T120 phosphorylation in HeLa cells, whereas hBUB1 is upregulated during mitosis, which corresponds with H2A T120 phosphorylation...
August 15, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Shoya Yasuda, Jiping Sun, Yang Zhou, Yanqing Wang, Qing Lu, Masayuki Yamamura, Ji-Yang Wang
The B cell receptor (BCR) transmits a tonic survival signal in the absence of antigen stimulation and an antigen-triggered survival signal. Mature B cells express two types of BCR, IgM and IgD, but it remains unclear how B cell survival is differentially regulated by these two receptors. We found that while crosslinking IgM on spleen B cells greatly enhanced their survival, crosslinking IgD did not enhance, but rather decreased, their survival. Consistently, crosslinking both IgM and IgD only moderately enhanced B cell survival, suggesting that IgM and IgD play opposing roles in B cell survival induced by BCR stimulation...
August 9, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Takeya Nakagawa, Mitsuhiro Yoneda, Miki Higashi, Yoshiaki Ohkuma, Takashi Ito
Regulation of the expression of diverse genes is essential for making possible the complexity of higher organisms, and the temporal and spatial regulation of gene expression allows for the alteration of cell types and growth patterns. A critical component of this regulation is the DNA sequence-specific binding of transcription factors (TFs). However, most TFs do not independently participate in gene transcriptional regulation, because they lack an effector function. Instead, TFs are thought to work by recruiting cofactors, including Mediator complex (Mediator), chromatin-remodeling complexes (CRCs), and histone-modifying complexes (HMCs)...
August 9, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Hidenobu Miyazawa, Masamichi Yamamoto, Yoshifumi Yamaguchi, Masayuki Miura
Developing embryos rewire energy metabolism for developmental processes. However, little is known about how metabolic rewiring is coupled with development in a spatiotemporal manner. Here, we show that mammalian embryos display plasticity of glucose metabolism in response to the extracellular environment at the neural tube closure (NTC) stage, when the intrauterine environment changes upon placentation. To study how embryos modulate their metabolic state upon environmental change, we analyzed the steady-state level of ATP upon exposure to extrauterine environments using both an enzymatic assay and a genetically encoded ATP sensor...
August 8, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Kenji Kitajima, Mai Kanokoda, Marino Nakajima, Takahiko Hara
The generation of mouse hematopoietic stem cells from hemogenic endothelial cells (HECs) in the aorta/gonad/mesonephros region of developing embryos requires a zinc finger transcription factor Gata2. In the previous study, an enforced expression of Gata2 in vitro promoted the production of HECs from mesodermal cells differentiated from mouse embryonic stem cells (ESCs). Our research group has previously demonstrated that the enforced expression of Gata2 in ESC-derived HECs enhances erythroid and megakaryocyte differentiation and inhibits macrophage differentiation...
August 8, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Ai Yamaguchi, Muneto Tatsumoto, Ritsuko Matsumura, Takuyuki Endo, Koichi Hirata, Isao Tokuda, Makoto Akashi
Almost all organisms maintain a circadian clock from birth to death to synchronize their own physiology and behavior with the earth's rotation. However, extensive studies based on animal experiments have showed that aging results in circadian dysfunction. Human studies have also indicated age-associated abnormal phase, reduced amplitude and enhanced fragmentation in circadian physiology and behavior, thereby strongly implying age-related dysfunction of the clock machinery. Here, we carried out functional assessment of the circadian clock machinery in elderly patients aged 83-94 with severe dementia who showed abnormal circadian behavior...
August 7, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Changkeun Kang, Kayoko Saso, Kazushige Ota, Masahito Kawazu, Takeshi Ueda, Hitoshi Okada
Obesity is a serious global health issue; however, the roles of genetics and epigenetics in the onset and progression of obesity are still not completely understood. The aim of this study was to determine the role of Kdm4b, which belongs to a subfamily of histone demethylases, in adipogenesis and fat metabolism in vivo. We established conditional Kdm4b knockout mice. Inactivation of Kdm4b in adipocytes (K4bKO) induced profound obesity in mice on a high fat diet (HFD). The HFD-fed K4bKO mice exhibited an increased volume of fat mass and higher expression levels of adipogenesis-related genes...
August 2, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Hitoshi Inada, Keiko Numayama-Tsuruta, Kentaro Mochizuki, Makiko Sasaki-Hoshino, Noriko Osumi
Fabp7 gene encodes a brain-specific fatty acid-binding protein that is widely used as a marker for neural stem cells. Here, we report that the activity of rat Fabp7 promoter was regulated directly by a transcription factor, Pax6. Deletion analyses identified an essential region (-837 to -64 from transcription start site) in the rat Fabp7 promoter. This region controls promoter activity in rat embryos and in the mouse cultured cell line MEB5. Over-expressing wild-type Pax6 or a dominant-negative Pax6 mutant enhanced and suppressed, respectively, the promoter activity...
July 9, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Mitsuhide Noshiro, Takeshi Kawamoto, Ayumu Nakashima, Noritsugu Ozaki, Toshinori Ueno, Masayumi Saeki, Kiyomasa Honda, Katsumi Fujimoto, Yukio Kato
Obesity is a major public health problem in developed countries resulting from increased food intake and decreased energy consumption and usually associated with abnormal lipid metabolism. Here, we show that DEC1, a basic helix-loop-helix transcription factor, plays an important role in the regulation of lipid consumption in mouse brown adipose tissue (BAT), which is the major site of thermogenesis. Homozygous Dec1 deletion attenuated high-fat-diet-induced obesity, adipocyte hypertrophy, fat volume and hepatic steatosis...
July 3, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
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