Cell structure and function最新文献_第3页

Sphingomyelin metabolism underlies Ras excitability for efficient cell migration and chemotaxis. 鞘磷脂代谢是 Ras 兴奋性的基础，可促进细胞高效迁移和趋化。

IF 2 4区生物学

Cell structure and function Pub Date : 2023-08-31 Epub Date: 2023-07-12 DOI: 10.1247/csf.23045

Da Young Shin, Hiroaki Takagi, Michio Hiroshima, Satomi Matsuoka, Masahiro Ueda

{"title":"Sphingomyelin metabolism underlies Ras excitability for efficient cell migration and chemotaxis.","authors":"Da Young Shin, Hiroaki Takagi, Michio Hiroshima, Satomi Matsuoka, Masahiro Ueda","doi":"10.1247/csf.23045","DOIUrl":"10.1247/csf.23045","url":null,"abstract":"In eukaryotic motile cells, the active Ras (Ras-GTP)-enriched domain is generated in an asymmetric manner on the cell membrane through the excitable dynamics of an intracellular signaling network. This asymmetric Ras signaling regulates pseudopod formation for both spontaneous random migration and chemoattractant-induced directional migration. While membrane lipids, such as sphingomyelin and phosphatidylserine, contribute to Ras signaling in various cell types, whether they are involved in the Ras excitability for cell motility is unknown. Here we report that functional Ras excitability requires the normal metabolism of sphingomyelin for efficient cell motility and chemotaxis. The pharmacological blockade of sphingomyelin metabolism by an acid-sphingomyelinase inhibitor, fendiline, and other inhibitors suppressed the excitable generation of the stable Ras-GTP-enriched domain. The suppressed excitability failed to invoke enough basal motility to achieve directed migration under shallow chemoattractant gradients. The fendiline-induced defects in Ras excitability, motility and stimulation-elicited directionality were due to an accumulation of sphingomyelin on the membrane, which could be recovered by exogenous sphingomyelinase or phosphatidylserine without changing the expression of Ras. These results indicate a novel regulatory mechanism of the excitable system by membrane lipids, in which sphingomyelin metabolism provides a membrane environment to ensure Ras excitation for efficient cellular motility and chemotaxis.Key words: cell polarity, cell migration, Ras, excitability, sphingomyelin.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"48 2","pages":"145-160"},"PeriodicalIF":2.0,"publicationDate":"2023-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11496829/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10498759","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

Two coral fluorescent proteins of distinct colors for sharp visualization of cell-cycle progression. 两种不同颜色的珊瑚荧光蛋白，可清晰显示细胞周期的进展。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2023-07-29 Epub Date: 2023-07-30 DOI: 10.1247/csf.23028

Ryoko Ando, Asako Sakaue-Sawano, Keiko Shoda, Atsushi Miyawaki

引用次数: 0

Proteomic analysis of fatty liver induced by starvation of medaka fish larvae. 青鳉幼鱼饥饿诱发脂肪肝的蛋白质组分析

IF 1.5 4区生物学

Cell structure and function Pub Date : 2023-07-19 Epub Date: 2023-06-28 DOI: 10.1247/csf.23014

Tomoyo Ikeda, Tokiro Ishikawa, Satoshi Ninagawa, Tetsuya Okada, Masaya Ono, Kazutoshi Mori

{"title":"Proteomic analysis of fatty liver induced by starvation of medaka fish larvae.","authors":"Tomoyo Ikeda, Tokiro Ishikawa, Satoshi Ninagawa, Tetsuya Okada, Masaya Ono, Kazutoshi Mori","doi":"10.1247/csf.23014","DOIUrl":"10.1247/csf.23014","url":null,"abstract":"When medaka fish (Oryzias latipes) larvae are grown in the absence of exogenous nutrition, the liver becomes dark and positive to Oil Red O staining from 7 days post-hatch (dph). We determined the mechanism of this starvation-induced development of fatty liver by proteomic analysis using livers obtained from larvae grown in the presence or absence of 2% glucose at 5 dph. Results showed that changes in the expression levels of enzymes involved in glycolysis or the tricarboxylic acid cycle were modest, whereas the expression levels of enzymes involved in amino acid catabolism or β-oxidation of fatty acids were significantly elevated, suggesting that they become major energy sources under starvation conditions. Expression levels of enzymes for the uptake and β-oxidation of fatty acids as well as synthesis of triacylglycerol were elevated, whereas those for the synthesis of cholesterol as well as export of cholesterol and triacylglycerol were decreased under starvation conditions, which explains the accumulation of triacylglycerol in the liver. Our results provide the basis for future research to understand how gene malfunction(s) affects the development of fatty liver, which can lead to nonalcoholic steatohepatitis and then to liver cirrhosis.Key words: amino acid catabolism, β-oxidation, triacylglycerol, cholesterol, export.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"48 2","pages":"123-133"},"PeriodicalIF":1.5,"publicationDate":"2023-07-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10915113/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9842855","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

Zebrafish imaging reveals hidden oncogenic-normal cell communication during primary tumorigenesis. 斑马鱼成像揭示了原发性肿瘤发生过程中隐藏的致癌细胞与正常细胞之间的交流。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2023-06-16 Epub Date: 2023-05-11 DOI: 10.1247/csf.23026

Yukinari Haraoka, Mai Miyake, Tohru Ishitani

引用次数: 0

Cell biology of protein-lipid conjugation. 蛋白质与脂质结合的细胞生物学。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2023-05-11 Epub Date: 2023-04-06 DOI: 10.1247/csf.23016

Jun-Ichi Sakamaki, Noboru Mizushima

引用次数: 1

A conserved WXXE motif is an apical delivery determinant of ABC transporter C subfamily isoforms. 保守的 WXXE 基序是 ABC 转运体 C 亚家族同种异构体的顶端输送决定因素。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2023-03-09 Epub Date: 2023-01-25 DOI: 10.1247/csf.22049

Md Shajedul Haque, Yoshikazu Emi, Masao Sakaguchi

{"title":"A conserved WXXE motif is an apical delivery determinant of ABC transporter C subfamily isoforms.","authors":"Md Shajedul Haque, Yoshikazu Emi, Masao Sakaguchi","doi":"10.1247/csf.22049","DOIUrl":"10.1247/csf.22049","url":null,"abstract":"ATP-binding cassette transporter isoform C7 (ABCC7), also designated as cystic fibrosis transmembrane conductance regulator (CFTR), is exclusively targeted to the apical plasma membrane of polarized epithelial cells. Although the apical localization of ABCC7 in epithelia is crucial for the Cl- excretion into lumens, the mechanism regulating its apical localization is poorly understood. In the present study, an apical localization determinant was identified in the N-terminal 80-amino acid long cytoplasmic region of ABCC7 (NT80). In HepG2 cells, overexpression of NT80 significantly disturbed the apical expression of ABCC7 in a competitive manner, suggesting the presence of a sorting determinant in this region. Deletion analysis identified a potential sorting information within a 20-amino acid long peptide (aa 41-60) of NT80. Alanine scanning mutagenesis of this region in full-length ABCC7 further narrowed down the apical localization determinant to four amino acids, W57DRE60. This WDRE sequence was conserved among vertebrate ABCC7 orthologs. Site-directed mutagenesis showed that W57 and E60 were critical for the apical expression of ABCC7, confirming a novel apical sorting determinant of ABCC7. Furthermore, a WXXE motif (tryptophan and glutamic acid residues with two-amino acid spacing) was found to be conserved among the N-terminal regions of apically localized ABCC members with 12-TM configuration. The significance of the WXXE motif was demonstrated for proper trafficking of ABCC4 to the apical plasma membrane.Key words: apical plasma membrane, sorting, ATP-binding cassette transporter, CFTR, MRP4.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"48 1","pages":"71-82"},"PeriodicalIF":1.5,"publicationDate":"2023-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10721954/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9135259","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

A non-nucleotide agonist that binds covalently to cysteine residues of STING. 一种非核苷酸激动剂，能与 STING 的半胱氨酸残基共价结合。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2023-02-16 Epub Date: 2022-12-28 DOI: 10.1247/csf.22085

Kentaro Matsumoto, Shenwei Ni, Hiroyuki Arai, Takashi Toyama, Yoshiro Saito, Takehiro Suzuki, Naoshi Dohmae, Kojiro Mukai, Tomohiko Taguchi

{"title":"A non-nucleotide agonist that binds covalently to cysteine residues of STING.","authors":"Kentaro Matsumoto, Shenwei Ni, Hiroyuki Arai, Takashi Toyama, Yoshiro Saito, Takehiro Suzuki, Naoshi Dohmae, Kojiro Mukai, Tomohiko Taguchi","doi":"10.1247/csf.22085","DOIUrl":"10.1247/csf.22085","url":null,"abstract":"Stimulator of interferon genes (STING) is an ER-localized transmembrane protein and the receptor for 2',3'-cyclic guanosine monophosphate-adenosine monophosphate (cGAMP), which is a second messenger produced by cGAMP synthase (cGAS), a cytosolic double-stranded DNA sensor. The cGAS-STING pathway plays a critical role in the innate immune response to infection of a variety of DNA pathogens through the induction of the type I interferons. Pharmacological activation of STING is a promising therapeutic strategy for cancer, thus the development of potent and selective STING agonists has been pursued. Here we report that mouse STING can be activated by phenylarsine oxide (PAO), a membrane permeable trivalent arsenic compound that preferentially reacts with thiol group of cysteine residue (Cys). The activation of STING with PAO does not require cGAS or cGAMP. Mass spectrometric analysis of the peptides generated by trypsin and chymotrypsin digestion of STING identifies several PAO adducts, suggesting that PAO covalently binds to STING. Screening of STING variants with single Cys to serine residues (Ser) reveals that Cys88 and Cys291 are critical to the response to PAO. STING activation with PAO, as with cGAMP, requires the ER-to-Golgi traffic and palmitoylation of STING. Our results identify a non-nucleotide STING agonist that does not target the cGAMP-binding pocket, and demonstrate that Cys of STING can be a novel target for the development of STING agonist.Key words: STING agonist, cysteine modification, innate immunity, phenylarsine oxide.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"48 1","pages":"59-70"},"PeriodicalIF":1.5,"publicationDate":"2023-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10721953/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10736938","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

Suppression of P-cadherin expression as a key regulatory element for embryonic stem cell stemness. 抑制 P-cadherin 的表达是胚胎干细胞干性的关键调控因素。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2023-02-08 Epub Date: 2022-12-28 DOI: 10.1247/csf.22060

Yuka Takeda, Shuji Matsuguchi, Sae Nozaki, Taisei Mihara, Junya Abe, Yohei Hirai

{"title":"Suppression of P-cadherin expression as a key regulatory element for embryonic stem cell stemness.","authors":"Yuka Takeda, Shuji Matsuguchi, Sae Nozaki, Taisei Mihara, Junya Abe, Yohei Hirai","doi":"10.1247/csf.22060","DOIUrl":"10.1247/csf.22060","url":null,"abstract":"In embryonic stem (ES) cell colonies, a small subpopulation that changes cell shape and loses pluripotency often appears in two-dimensional (2D) cultures, even in the presence of a stemness factor. We have previously shown that membrane translocation of the syntaxin4, t-SNARE protein contributes to this phenomenon. Here, we show that ES cells in three-dimensional (3D) aggregates do not succumb to extruded syntaxin4 owing to suppressed expression of P-cadherin protein. While extracellular expression of syntaxin4 led to the striking upregulation of P-cadherin mRNA in both 2D and 3D-ES cells, morphological changes and appreciable expression of P-cadherin protein were detected only in 2D-ES cells. Importantly, the introduction of an expression cassette for P-cadherin practically reproduced the effects induced by extracellular syntaxin4, where the transgene product was clearly detected in 2D-, but not 3D-ES cells. An expression construct for P-cadherin-Venus harboring an in-frame insertion of the P2A sequence at the joint region gave fluorescent signals only in the cytoplasm of 2D-ES cells, demonstrating translational regulation of P-cadherin. These results provide the mechanistic insight into the uncontrollable differentiation in 2D-ES cells and shed light on the validity of the \"embryoid body protocol commonly used for ES cell handling\" for directional differentiation.Key words: differentiation, embryoid body, ES cells, P-cadherin, syntaxin4.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"48 1","pages":"49-57"},"PeriodicalIF":1.5,"publicationDate":"2023-02-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10721948/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10680262","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

Enhancement of Vivid-based photo-activatable Gal4 transcription factor in mammalian cells. 增强哺乳动物细胞中基于 Vivid 的可光激活 Gal4 转录因子。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2023-02-08 Epub Date: 2022-12-16 DOI: 10.1247/csf.22074

Shinji C Nagasaki, Tomonori D Fukuda, Mayumi Yamada, Yusuke Iii Suzuki, Ryo Kakutani, Adam T Guy, Itaru Imayoshi

{"title":"Enhancement of Vivid-based photo-activatable Gal4 transcription factor in mammalian cells.","authors":"Shinji C Nagasaki, Tomonori D Fukuda, Mayumi Yamada, Yusuke Iii Suzuki, Ryo Kakutani, Adam T Guy, Itaru Imayoshi","doi":"10.1247/csf.22074","DOIUrl":"10.1247/csf.22074","url":null,"abstract":"The Gal4/UAS system is a versatile tool to manipulate exogenous gene expression of cells spatially and temporally in many model organisms. Many variations of light-controllable Gal4/UAS system are now available, following the development of photo-activatable (PA) molecular switches and integration of these tools. However, many PA-Gal4 transcription factors have undesired background transcription activities even in dark conditions, and this severely attenuates reliable light-controlled gene expression. Therefore, it is important to develop reliable PA-Gal4 transcription factors with robust light-induced gene expression and limited background activity. By optimization of synthetic PA-Gal4 transcription factors, we have validated configurations of Gal4 DNA biding domain, transcription activation domain and blue light-dependent dimer formation molecule Vivid (VVD), and applied types of transcription activation domains to develop a new PA-Gal4 transcription factor we have named eGAV (enhanced Gal4-VVD transcription factor). Background activity of eGAV in dark conditions was significantly lower than that of hGAVPO, a commonly used PA-Gal4 transcription factor, and maximum light-induced gene expression levels were also improved. Light-controlled gene expression was verified in cultured HEK293T cells with plasmid-transient transfections, and in mouse EpH4 cells with lentivirus vector-mediated transduction. Furthermore, light-controlled eGAV-mediated transcription was confirmed in transfected neural stem cells and progenitors in developing and adult mouse brain and chick spinal cord, and in adult mouse hepatocytes, demonstrating that eGAV can be applied to a wide range of experimental systems and model organisms.Key words: optogenetics, Gal4/UAS system, transcription, gene expression, Vivid.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"48 1","pages":"31-47"},"PeriodicalIF":1.5,"publicationDate":"2023-02-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10721950/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9324635","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

Role of phosphatidylserine in the localization of cell surface membrane proteins in yeast. 磷脂酰丝氨酸在酵母细胞表面膜蛋白定位中的作用

IF 1.5 4区生物学

Cell structure and function Pub Date : 2023-01-28 Epub Date: 2022-12-15 DOI: 10.1247/csf.22081

Ryutaro Kashikuma, Makoto Nagano, Hiroki Shimamura, Kouya Nukaga, Ikumi Katsumata, Junko Y Toshima, Jiro Toshima

{"title":"Role of phosphatidylserine in the localization of cell surface membrane proteins in yeast.","authors":"Ryutaro Kashikuma, Makoto Nagano, Hiroki Shimamura, Kouya Nukaga, Ikumi Katsumata, Junko Y Toshima, Jiro Toshima","doi":"10.1247/csf.22081","DOIUrl":"10.1247/csf.22081","url":null,"abstract":"Phosphatidylserine (PS) is a constituent of the cell membrane, being especially abundant in the cytoplasmic leaflet, and plays important roles in a number of cellular functions, including the formation of cell polarity and intracellular vesicle transport. Several studies in mammalian cells have suggested the role of PS in retrograde membrane traffic through endosomes, but in yeast, where PS is localized primarily at the plasma membrane (PM), the role in intracellular organelles remains unclear. Additionally, it is reported that polarized endocytic site formation is defective in PS-depleted yeast cells, but the role in the endocytic machinery has not been well understood. In this study, to clarify the role of PS in the endocytic pathway, we analyzed the effect of PS depletion on endocytic internalization and post-endocytic transport. We demonstrated that in cell lacking the PS synthase Cho1p (cho1Δ cell), binding and internalization of mating pheromone α-factor into the cell was severely impaired. Interestingly, the processes of endocytosis were mostly unaffected, but protein transport from the trans-Golgi network (TGN) to the PM was defective and localization of cell surface proteins was severely impaired in cho1Δ cells. We also showed that PS accumulated in intracellular compartments in cells lacking Rcy1p and Vps52p, both of which are implicated in endosome-to-PM transport via the TGN, and that the number of Snx4p-residing endosomes was increased in cho1Δ cells. These results suggest that PS plays a crucial role in the transport and localization of cell surface membrane proteins.Key words: phosphatidylserine, endocytosis, recycling, vesicle transport.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"48 1","pages":"19-30"},"PeriodicalIF":1.5,"publicationDate":"2023-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10725852/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10655171","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