Cell structure and function最新文献_第5页

A Dual Promoter System to Monitor IFN-γ Signaling in vivo at Single-cell Resolution. 以单细胞分辨率监测体内 IFN-γ 信号的双启动子系统

IF 1.5 4区生物学

Cell structure and function Pub Date : 2021-12-22 Epub Date: 2021-11-06 DOI: 10.1247/csf.21052

Taisei Tanaka, Yoshinobu Konishi, Hiroshi Ichise, Shinya Tsukiji, Michiyuki Matsuda, Kenta Terai

{"title":"A Dual Promoter System to Monitor IFN-γ Signaling in vivo at Single-cell Resolution.","authors":"Taisei Tanaka, Yoshinobu Konishi, Hiroshi Ichise, Shinya Tsukiji, Michiyuki Matsuda, Kenta Terai","doi":"10.1247/csf.21052","DOIUrl":"10.1247/csf.21052","url":null,"abstract":"IFN-γ secreted from immune cells exerts pleiotropic effects on tumor cells, including induction of immune checkpoint and antigen presentation, growth inhibition, and apoptosis induction. We combined a dual promoter system with an IFN-γ signaling responsive promoter to generate a reporter named the interferon sensing probe (ISP), which quantitates the response to IFN-γ by means of fluorescence and bioluminescence. The integration site effect of the transgene is compensated for by the PGK promoter-driven expression of a fluorescent protein. Among five potential IFN-γ-responsive elements, we found that the interferon γ-activated sequence (GAS) exhibited the best performance. When ISP-GAS was introduced into four cell lines and subjected to IFN-γ stimulation, dose-dependency was observed with an EC50 ranging from 0.2 to 0.9 ng/mL, indicating that ISP-GAS can be generally used as a sensitive biosensor of IFN-γ response. In a syngeneic transplantation model, the ISP-GAS-expressing cancer cells exhibited bioluminescence and fluorescence signals in an IFN-γ receptor-dependent manner. Thus, ISP-GAS could be used to quantitatively monitor the IFN-γ response both in vitro and in vivo.Key words: in vivo imaging, tumor microenvironment, interferon-gamma, dual promoter system.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"46 2","pages":"103-111"},"PeriodicalIF":1.5,"publicationDate":"2021-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10511040/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39596761","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Visualization of DNA Replication in Single Chromosome by Stable Isotope Labeling. 通过稳定同位素标记观察单个染色体中的 DNA 复制。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2021-11-20 Epub Date: 2021-09-25 DOI: 10.1247/csf.21011

Kosuke Nagata, Ken-Ichi Bajo, Hideyuki Mitomo, Ryosuke Fujita, Ryota Uehara, Kuniharu Ijiro, Hisayoshi Yurimoto

引用次数: 0

Nuclear RNA Regulation by XRN2 and XTBD Family Proteins. XRN2 和 XTBD 家族蛋白对核 RNA 的调控

IF 1.5 4区生物学

Cell structure and function Pub Date : 2021-11-06 Epub Date: 2021-09-03 DOI: 10.1247/csf.21041

Ilkin Aygün, Takashi S Miki

引用次数: 0

In vivo Roles of Rab27 and Its Effectors in Exocytosis. Rab27 及其效应因子在体内外吞过程中的作用

IF 1.5 4区生物学

Cell structure and function Pub Date : 2021-11-06 Epub Date: 2021-09-04 DOI: 10.1247/csf.21043

Tetsuro Izumi

{"title":"In vivo Roles of Rab27 and Its Effectors in Exocytosis.","authors":"Tetsuro Izumi","doi":"10.1247/csf.21043","DOIUrl":"10.1247/csf.21043","url":null,"abstract":"The monomeric GTPase Rab27 regulates exocytosis of a broad range of vesicles in multicellular organisms. Several effectors bind GTP-bound Rab27a and/or Rab27b on secretory vesicles to execute a series of exocytic steps, such as vesicle maturation, movement along microtubules, anchoring within the peripheral F-actin network, and tethering to the plasma membrane, via interactions with specific proteins and membrane lipids in a local milieu. Although Rab27 effectors generally promote exocytosis, they can also temporarily restrict it when they are involved in the rate-limiting step. Genetic alterations in Rab27-related molecules cause discrete diseases manifesting pigment dilution and immunodeficiency, and can also affect common diseases such as diabetes and cancer in complex ways. Although the function and mechanism of action of these effectors have been explored, it is unclear how multiple effectors act in coordination within a cell to regulate the secretory process as a whole. It seems that Rab27 and various effectors constitutively reside on individual vesicles to perform consecutive exocytic steps. The present review describes the unique properties and in vivo roles of the Rab27 system, and the functional relationship among different effectors coexpressed in single cells, with pancreatic beta cells used as an example.Key words: membrane trafficking, regulated exocytosis, insulin granules, pancreatic beta cells.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"46 2","pages":"79-94"},"PeriodicalIF":1.5,"publicationDate":"2021-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10511049/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39384597","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Hrd1-dependent Degradation of the Unassembled PIGK Subunit of the GPI Transamidase Complex. 依赖于 Hrd1 的 GPI 跨酰胺酶复合物未组装 PIGK 亚基降解。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2021-09-03 Epub Date: 2021-06-30 DOI: 10.1247/csf.21019

Kohei Kawaguchi, Miki Yamamoto-Hino, Yoshiko Murakami, Taroh Kinoshita, Satoshi Goto

引用次数: 1

Expression Patterns and Levels of All Tubulin Isotypes Analyzed in GFP Knock-In C. elegans Strains. 在 GFP 基因敲入 C. elegans 株系中分析的所有微管蛋白异型的表达模式和水平。

IF 2 4区生物学

Cell structure and function Pub Date : 2021-06-30 Epub Date: 2021-05-08 DOI: 10.1247/csf.21022

Kei Nishida, Kenta Tsuchiya, Hiroyuki Obinata, Shizuka Onodera, Yu Honda, Yen-Cheng Lai, Nami Haruta, Asako Sugimoto

{"title":"Expression Patterns and Levels of All Tubulin Isotypes Analyzed in GFP Knock-In C. elegans Strains.","authors":"Kei Nishida, Kenta Tsuchiya, Hiroyuki Obinata, Shizuka Onodera, Yu Honda, Yen-Cheng Lai, Nami Haruta, Asako Sugimoto","doi":"10.1247/csf.21022","DOIUrl":"10.1247/csf.21022","url":null,"abstract":"Most organisms have multiple α- and β-tubulin isotypes that likely contribute to the diversity of microtubule (MT) functions. To understand the functional differences of tubulin isotypes in Caenorhabditis elegans, which has nine α-tubulin isotypes and six β-tubulin isotypes, we systematically constructed null mutants and GFP-fusion strains for all tubulin isotypes with the CRISPR/Cas9 system and analyzed their expression patterns and levels in adult hermaphrodites. Four isotypes-α-tubulins TBA-1 and TBA-2 and β-tubulins TBB-1 and TBB-2-were expressed in virtually all tissues, with a distinct tissue-specific spectrum. Other isotypes were expressed in specific tissues or cell types at significantly lower levels than the broadly expressed isotypes. Four isotypes (TBA-5, TBA-6, TBA-9, and TBB-4) were expressed in different subsets of ciliated sensory neurons, and TBB-4 was inefficiently incorporated into mitotic spindle MTs. Taken together, we propose that MTs in C. elegans are mainly composed of four broadly expressed tubulin isotypes and that incorporation of a small amount of tissue-specific isotypes may contribute to tissue-specific MT properties. These newly constructed strains will be useful for further elucidating the distinct roles of tubulin isotypes.Key words: tubulin isotypes, microtubules, C. elegans.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"46 1","pages":"51-64"},"PeriodicalIF":2.0,"publicationDate":"2021-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10511039/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38883723","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Multiple Ways for Stress Sensing and Regulation of the Endoplasmic Reticulum-stress Sensors. 内质网应力传感器的多种应力感应和调节方式

IF 2 4区生物学

Cell structure and function Pub Date : 2021-05-22 Epub Date: 2021-03-26 DOI: 10.1247/csf.21015

Quynh Giang Le, Yukio Kimata

{"title":"Multiple Ways for Stress Sensing and Regulation of the Endoplasmic Reticulum-stress Sensors.","authors":"Quynh Giang Le, Yukio Kimata","doi":"10.1247/csf.21015","DOIUrl":"10.1247/csf.21015","url":null,"abstract":"Dysfunction of the endoplasmic reticulum (ER), so-called ER stress, is accompanied with accumulation of unfolded proteins in the ER. Eukaryotic cells commonly have an ER-located transmembrane protein, Ire1, which triggers cellular protective events against ER stress. In animal cells, PERK and ATF6 also initiate the ER-stress response. As a common strategy to control the activity of these ER-stress sensors, an ER-resident molecular chaperone, BiP, serves as their negative regulator, and dissociates from them in response to ER stress. Although it sounds reasonable that unfolded proteins and Ire1 compete for BiP association, some publications argue against this competition model. Moreover, yeast Ire1 (and possibly also the mammalian major Ire1 paralogue IRE1α) directly detects ER-accumulated unfolded proteins, and subsequently oligomerizes for its further activation. Apart from protein misfolding, the saturation of membrane phospholipids is another outcome of ER-stressing stimuli, which is sensed by the transmembrane domain of Ire1. This review describes the canonical and up-to-date insights concerning stress-sensing and regulatory mechanisms of yeast Ire1 and metazoan ER-stress sensors.Key words: endoplasmic reticulum, stress, unfolded protein response, molecular chaperone.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"46 1","pages":"37-49"},"PeriodicalIF":2.0,"publicationDate":"2021-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10511038/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25537938","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Dispensability of Tubulin Acetylation for 15-protofilament Microtubule Formation in the Mammalian Cochlea. 哺乳动物耳蜗中 15-原丝微管形成过程中微管蛋白乙酰化的不可或缺性

IF 2 4区生物学

Cell structure and function Pub Date : 2021-03-16 Epub Date: 2021-02-06 DOI: 10.1247/csf.20057

Justine Renauld, Nicolas Thelen, Odile Bartholomé, Brigitte Malgrange, Marc Thiry

引用次数: 0

Knock-in of Labeled Proteins into 5'UTR Enables Highly Efficient Generation of Stable Cell Lines. 将标记蛋白质敲入 5'UTR 可高效生成稳定的细胞系。

IF 2 4区生物学

Cell structure and function Pub Date : 2021-03-16 Epub Date: 2021-01-26 DOI: 10.1247/csf.21002

Faryal Ijaz, Koji Ikegami

{"title":"Knock-in of Labeled Proteins into 5'UTR Enables Highly Efficient Generation of Stable Cell Lines.","authors":"Faryal Ijaz, Koji Ikegami","doi":"10.1247/csf.21002","DOIUrl":"10.1247/csf.21002","url":null,"abstract":"Stable cell lines and animal models expressing tagged proteins are important tools for studying behaviors of cells and molecules. Several molecular biology technologies have been applied with varying degrees of success and efficiencies to establish cell lines expressing tagged proteins. Here we applied CRISPR/Cas9 for the knock-in of tagged proteins into the 5'UTR of the endogenous gene loci. With this 5'UTR-targeting knock-in strategy, stable cell lines expressing Arl13b-Venus, Reep6-HA, and EGFP-alpha-tubulin were established with high efficiencies ranging from 50 to 80% in antibiotic selected cells. The localization of the knock-in proteins were identical to that of the endogenous proteins in wild-type cells and showed homogenous expression. Moreover, the expression of knock-in EGFP-alpha-tubulin from the endogenous promoter was stable over long-term culture. We further demonstrated that the fluorescent signals were enough for a long time time-lapse imaging. The fluorescent signals were distinctly visible during the whole duration of the time-lapse imaging and showed specific subcellular localizations. Altogether, our strategy demonstrates that 5'UTR is an amenable site to generate cell lines for the stable expression of tagged proteins from endogenous loci in mammalian cells.Key words: CRISPR/Cas9, knock-in, primary cilium, UTR, tubulin.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"46 1","pages":"21-35"},"PeriodicalIF":2.0,"publicationDate":"2021-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10511053/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38867755","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Mevalonate Pathway-mediated ER Homeostasis Is Required for Haploid Stability in Human Somatic Cells. 人类体细胞单倍体稳定性需要甲羟戊酸途径介导的ER平衡

IF 2 4区生物学

Cell structure and function Pub Date : 2021-02-19 Epub Date: 2020-12-22 DOI: 10.1247/csf.20055

Kan Yaguchi, Kimino Sato, Koya Yoshizawa, Daisuke Mikami, Kohei Yuyama, Yasuyuki Igarashi, Gabor Banhegyi, Eva Margittai, Ryota Uehara

{"title":"Mevalonate Pathway-mediated ER Homeostasis Is Required for Haploid Stability in Human Somatic Cells.","authors":"Kan Yaguchi, Kimino Sato, Koya Yoshizawa, Daisuke Mikami, Kohei Yuyama, Yasuyuki Igarashi, Gabor Banhegyi, Eva Margittai, Ryota Uehara","doi":"10.1247/csf.20055","DOIUrl":"10.1247/csf.20055","url":null,"abstract":"The somatic haploidy is unstable in diplontic animals, but cellular processes determining haploid stability remain elusive. Here, we found that inhibition of mevalonate pathway by pitavastatin, a widely used cholesterol-lowering drug, drastically destabilized the haploid state in HAP1 cells. Interestingly, cholesterol supplementation did not restore haploid stability in pitavastatin-treated cells, and cholesterol inhibitor U18666A did not phenocopy haploid destabilization. These results ruled out the involvement of cholesterol in haploid stability. Besides cholesterol perturbation, pitavastatin induced endoplasmic reticulum (ER) stress, the suppression of which by a chemical chaperon significantly restored haploid stability in pitavastatin-treated cells. Our data demonstrate the involvement of the mevalonate pathway in the stability of the haploid state in human somatic cells through managing ER stress, highlighting a novel link between ploidy and ER homeostatic control.Key words: haploid, ER stress, Mevalonate pathway.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"46 1","pages":"1-9"},"PeriodicalIF":2.0,"publicationDate":"2021-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10511059/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38749324","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0