Cell structure and function最新文献

筛选
英文 中文
A sensitive ERK fluorescent probe reveals the significance of minimal EGF-induced transcription.
IF 2 4区 生物学
Cell structure and function Pub Date : 2024-12-18 DOI: 10.1247/csf.24070
Zhang Weisheng, Jun Nakayama, Yukino Inomata, Shigeki Higashiyama, Toru Hiratsuka
{"title":"A sensitive ERK fluorescent probe reveals the significance of minimal EGF-induced transcription.","authors":"Zhang Weisheng, Jun Nakayama, Yukino Inomata, Shigeki Higashiyama, Toru Hiratsuka","doi":"10.1247/csf.24070","DOIUrl":"https://doi.org/10.1247/csf.24070","url":null,"abstract":"<p><p>Extracellular signal-regulated kinase (ERK) regulates multiple cellular functions through distinct activation patterns. Genetically encoded fluorescent probes are instrumental in dissecting the ERK activity dynamics in living cells. Here we modified a previously reported Förster resonance energy transfer (FRET) probe for ERK, EKAREN5 by replacing its mTurquoise2 and YPet sequences with mTurquoise-GL and a synonymous codon variant of YPet, respectively. The modified biosensor, EKAREN5-gl showed an increased sensitivity to EGF-induced ERK activation responding to a very low dose (20 pg/ml) of EGF stimulation. We quantitatively characterized two FRET-based ERK probes, EKAREN5 and EKAREN5-gl, and a subcellular kinase translocation-based probe, ERK-KTR. We found the three biosensors differently respond to EGF stimulations with different intensity, duration, and latency. Furthermore, we investigated how the minimal EGF-induced ERK activation affects the downstream transcription in HeLa cells by comprehensive transcriptional analysis. We found the minimal ERK activation leads to a distinct transcriptional pattern from those induced by higher ERK activations. Our study highlights the significance of sensitive fluorescent probes to understand cellular signal dynamics and the role of minimal ERK activation in regulating transcription.Key words: fluorescent probe, ERK, FRET, KTR.</p>","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":" ","pages":""},"PeriodicalIF":2.0,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142853189","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Possible roles of CAHS proteins from Tardigrade in osmotic stress tolerance in mammalian cells. CAHS蛋白在哺乳动物细胞的渗透胁迫耐受性中的可能作用。
IF 2 4区 生物学
Cell structure and function Pub Date : 2024-12-14 Epub Date: 2024-12-05 DOI: 10.1247/csf.24035
Takahiro Bino, Yuhei Goto, Gembu Maryu, Kazuharu Arakawa, Kazuhiro Aoki
{"title":"Possible roles of CAHS proteins from Tardigrade in osmotic stress tolerance in mammalian cells.","authors":"Takahiro Bino, Yuhei Goto, Gembu Maryu, Kazuharu Arakawa, Kazuhiro Aoki","doi":"10.1247/csf.24035","DOIUrl":"10.1247/csf.24035","url":null,"abstract":"<p><p>Anhydrobiosis, a phenomenon in which organisms survive extreme dehydration by entering a reversible ametabolic state, is a remarkable example of survival strategies. This study focuses on anhydrobiosis in tardigrades, which are known for their resilience to severe environmental conditions. Tardigrades utilize several protective mechanisms against desiccation, notably the constitutive expression of cytoplasmic abundant heat soluble (CAHS) proteins in Ramazzottius varieornatus. These proteins share similarities in their amphiphatic alpha helices with late embryogenesis abundant (LEA) proteins, but differ significantly in their amino acid sequences. In this study, we further explored the functionality of CAHS proteins by analyzing their role in aggregation and tolerance to hyperosmotic stress in mammalian cells. Using live cell imaging, we examined the subcellular localization of several CAHS and LEA proteins in response to hyperosmotic stress. The expression of CAHS1, CAHS3, and CAHS8 tended to enhance the resilience to the hyperosmotic conditions. These findings not only deepen our understanding of the molecular mechanisms of anhydrobiosis but also highlight the potential of CAHS proteins as cryoprotectants.Key words: anhydrobiosis, Tardigrades, live imaging, disordered proteins, desiccation tolerance.</p>","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":" ","pages":"123-133"},"PeriodicalIF":2.0,"publicationDate":"2024-12-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142680980","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Multi-color fluorescence live-cell imaging in Dictyostelium discoideum.
IF 2 4区 生物学
Cell structure and function Pub Date : 2024-12-04 DOI: 10.1247/csf.24065
Hidenori Hashimura, Satoshi Kuwana, Hibiki Nakagwa, Kenichi Abe, Tomoko Adachi, Toyoko Sugita, Shoko Fujishiro, Gen Honda, Satoshi Sawai
{"title":"Multi-color fluorescence live-cell imaging in Dictyostelium discoideum.","authors":"Hidenori Hashimura, Satoshi Kuwana, Hibiki Nakagwa, Kenichi Abe, Tomoko Adachi, Toyoko Sugita, Shoko Fujishiro, Gen Honda, Satoshi Sawai","doi":"10.1247/csf.24065","DOIUrl":"https://doi.org/10.1247/csf.24065","url":null,"abstract":"<p><p>The cellular slime mold Dictyostelium discoideum, a member of the Amoebozoa, has been extensively studied in cell and developmental biology. D. discoideum is unique in that they are genetically tractable, with a wealth of data accumulated over half a century of research. Fluorescence live-cell imaging of D. discoideum has greatly facilitated studies on fundamental topics, including cytokinesis, phagocytosis, and cell migration. Additionally, its unique life cycle places Dictyostelium at the forefront of understanding aggregative multicellularity, a recurring evolutionary trait found across the Opisthokonta and Amoebozoa clades. The use of multiple fluorescent proteins (FP) and labels with separable spectral properties is critical for tracking cells in aggregates and identifying co-occurring biomolecular events and factors that underlie the dynamics of the cytoskeleton, membrane lipids, second messengers, and gene expression. However, in D. discoideum, the number of frequently used FP species is limited to two or three. In this study, we explored the use of new-generation FP for practical 4- to 5-color fluorescence imaging of D. discoideum. We showed that the yellow fluorescent protein Achilles and the red fluorescent protein mScarlet-I both yield high signals and allow sensitive detection of rapid gene induction. The color palette was further expanded to include blue (mTagBFP2 and mTurquosie2), large Stoke-shift LSSmGFP, and near-infrared (miRFP670nano3) FPs, in addition to the HaloTag ligand SaraFluor 650T. Thus, we demonstrated the feasibility of deploying 4- and 5- color imaging of D. discoideum using conventional confocal microscopy.Key words: fluorescence imaging, organelle, cytoskeleton, small GTPase, Dictyostelium.</p>","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":" ","pages":""},"PeriodicalIF":2.0,"publicationDate":"2024-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142778837","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
The expression of the formin Fhod3 in mouse tongue striated muscle. 形蛋白 Fhod3 在小鼠舌横纹肌中的表达。
IF 2 4区 生物学
Cell structure and function Pub Date : 2024-11-07 Epub Date: 2024-10-10 DOI: 10.1247/csf.24044
Hikaru Nakagawa, Yohko Kage, Ayako Miura, Hikmawan Wahyu Sulistomo, Sho Matsuyama, Yoshihiro Yamashita, Ryu Takeya
{"title":"The expression of the formin Fhod3 in mouse tongue striated muscle.","authors":"Hikaru Nakagawa, Yohko Kage, Ayako Miura, Hikmawan Wahyu Sulistomo, Sho Matsuyama, Yoshihiro Yamashita, Ryu Takeya","doi":"10.1247/csf.24044","DOIUrl":"10.1247/csf.24044","url":null,"abstract":"<p><p>The sarcomere is the contractile unit of striated muscle and is composed of actin and myosin filaments. There is increasing evidence to support that actin assembly mediated by Fhod3, a member of the formin family of proteins, is critical for sarcomere formation and maintenance in cardiac muscle. Fhod3, which is abundantly expressed in the heart, localizes to the center of sarcomeres and contributes to the regulation of the cardiac function, as evidenced by the fact that mutations in Fhod3 cause cardiomyopathy. However, the role of Fhod3 in skeletal muscle, another type of striated muscle, is unclear. We herein show that Fhod3 is expressed in the tongue at both mRNA and protein levels, although in smaller amounts than in the heart. To determine the physiological role of Fhod3 expressed in the tongue, we generated embryos lacking Fhod3 in the tongue. The tongue tissue of the Fhod3-depleted embryos did not show any significant structural defects, suggesting that Fhod3 is dispensable for normal development of the mouse tongue. Unexpectedly, the immunostaining analysis revealed the absence of specific sarcomeric signals for Fhod3 in the wild-type tongue when compared to the Fhod3-depleted tongue as a negative control, despite the use of antibodies that had previously been validated by immunostaining of heart tissues. Taken together, although Fhod3 protein is expressed at a significant level in the tongue, Fhod3 in the tongue does not appear to exhibit the same sarcomeric pattern as observed in the heart, suggesting a different role for Fhod3 in the tongue muscles.Key words: actin, formin, sarcomere, striated muscle.</p>","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":" ","pages":"111-122"},"PeriodicalIF":2.0,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142388446","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Cell-wide arrangement of Golgi/RE units depends on the microtubule organization. 高尔基体/RE 单元在整个细胞内的排列取决于微管组织。
IF 2 4区 生物学
Cell structure and function Pub Date : 2024-11-07 Epub Date: 2024-10-03 DOI: 10.1247/csf.24055
Tatsuya Tago, Syara Fujii, Shogo Sasaki, Maki Shirae-Kurabayashi, Naoaki Sakamoto, Takashi Yamamoto, Makoto Maeda, Tatsuya Ueki, Takunori Satoh, Akiko K Satoh
{"title":"Cell-wide arrangement of Golgi/RE units depends on the microtubule organization.","authors":"Tatsuya Tago, Syara Fujii, Shogo Sasaki, Maki Shirae-Kurabayashi, Naoaki Sakamoto, Takashi Yamamoto, Makoto Maeda, Tatsuya Ueki, Takunori Satoh, Akiko K Satoh","doi":"10.1247/csf.24055","DOIUrl":"10.1247/csf.24055","url":null,"abstract":"<p><p>We have previously shown that Golgi stacks and recycling endosomes (REs) exist as Golgi/RE units in sea urchin embryos. In this study, we showed that Golgi/RE units were scattered throughout the cytoplasm at early developmental stages but gathered to form a \"Golgi ring\" surrounding the centric REs at the blastula stage. This change in the cell-wide arrangement of Golgi/RE units coincided with a dramatic change in microtubule organization from a randomly oriented cortical pattern to radial arrays under the apical plasma membrane. A single gigantic Golgi apparatus surrounding centric RE is clearly associated with the center of the radial microtubule arrays. Furthermore, we found that in some animal species belonging to different clades, Golgi stacks lack lateral connections but are likely centralized by microtubule motors. These results suggest that Golgi centralization depends on the organization of the microtubule array in addition to the lateral linking between Golgi stacks.Key words: Golgi stack, recycling endosome, Golgi-ribbon, microtubule, cilium, sea urchin, ascidian.</p>","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":" ","pages":"101-110"},"PeriodicalIF":2.0,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142368182","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Molecular alterations associated with pathophysiology in liver-specific ZO-1 and ZO-2 knockout mice. 与肝特异性 ZO-1 和 ZO-2 基因敲除小鼠病理生理学相关的分子变化
IF 2 4区 生物学
Cell structure and function Pub Date : 2024-10-26 Epub Date: 2024-09-26 DOI: 10.1247/csf.24046
Masahiko Itoh, Kenji Watanabe, Yoichi Mizukami, Hiroyuki Sugimoto
{"title":"Molecular alterations associated with pathophysiology in liver-specific ZO-1 and ZO-2 knockout mice.","authors":"Masahiko Itoh, Kenji Watanabe, Yoichi Mizukami, Hiroyuki Sugimoto","doi":"10.1247/csf.24046","DOIUrl":"10.1247/csf.24046","url":null,"abstract":"<p><p>The liver is a complex organ with a highly organized structure in which tight junctions (TJs) play an important role in maintaining their function by regulating barrier properties and cellular polarity. Dysfunction of TJs is associated with liver diseases, including progressive familial intrahepatic cholestasis (PFIC). In this study, we investigated the molecular alterations in a liver-specific ZO-1 and ZO-2 double-knockout (DKO) mouse model, which exhibits features resembling those of PFIC4 patients with mutations in the ZO-2 gene. RNA-seq analysis revealed the upregulation of genes involved in the oxidative stress response, xenobiotic metabolism, and cholesterol metabolism in DKO livers. Conversely, the expression of genes regulated by HNF4α was lower in DKO livers than in the wild-type controls. Furthermore, age-associated analysis elucidated the timing and progression of these pathway changes as well as alterations in molecules related to TJs and apical polarity. Our research uncovered previously unknown implications of ZO-1 and ZO-2 in liver physiology and provides new insights into the molecular pathogenesis of PFIC4 and other tight junction-related liver diseases. These findings contribute to a better understanding of the complex mechanisms underlying liver function and dysfunction and may lead to the development of novel therapeutic strategies for liver diseases associated with tight junction impairment.Key words: tight junctions, ZO-1/ZO-2 knockout mouse, liver, transcriptome analysis, molecular pathological progression.</p>","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":" ","pages":"83-99"},"PeriodicalIF":2.0,"publicationDate":"2024-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142342647","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Visualization of ER-to-Golgi trafficking of procollagen X. 胶原蛋白 X 从 ER 到高尔基体运输的可视化。
IF 2 4区 生物学
Cell structure and function Pub Date : 2024-10-02 Epub Date: 2024-09-06 DOI: 10.1247/csf.24024
Yuan Ximin, Hitoshi Hashimoto, Ikuo Wada, Nobuko Hosokawa
{"title":"Visualization of ER-to-Golgi trafficking of procollagen X.","authors":"Yuan Ximin, Hitoshi Hashimoto, Ikuo Wada, Nobuko Hosokawa","doi":"10.1247/csf.24024","DOIUrl":"10.1247/csf.24024","url":null,"abstract":"<p><p>Collagen is the most abundant protein in the extracellular matrix of animals, and 28 types of collagen have been reported in humans. We previously analyzed the endoplasmic reticulum (ER)-to-Golgi transport of fibril-forming type III collagen (Hirata et al., 2022) and network-forming type IV collagen (Matsui et al., 2020), both of which have long collagenous triple-helical regions. To understand the ER-to-Golgi trafficking of various types of collagens, we analyzed the transport of short-chain type X collagen in this study. We fused cysteine-free GFP to the N-telopeptide region of procollagen X (GFP-COL10A1), as employed in our previous analysis of procollagens III and IV, and analyzed its transport by live-cell imaging. Procollagen X was transported to the Golgi apparatus via vesicular and tubular carriers containing ERGIC53 and RAB1B, similar to those used for procollagen III. Carriers containing procollagen X probably used the same transport processes as those containing conventional cargoes such as α<sub>1</sub>-antitrypsin. SAR1, TANGO1, SLY1/SCFD1, and BET3/TRAPPC3 were required for trafficking of procollagen X, which are different from the factors required for trafficking of procollagens III (SAR1, TANGO1, and CUL3) and IV (SAR1 and SLY1/SCFD1). These findings reveal that accommodation of various types of collagens with different shapes into carriers may require fine-tuning of the ER-to-Golgi transport machinery.Key words: collagen, GFP-procollagen X, ER-to-Golgi trafficking, export from ER, TANGO1.</p>","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":" ","pages":"67-81"},"PeriodicalIF":2.0,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142153233","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Deep learning-based segmentation of subcellular organelles in high-resolution phase-contrast images. 基于深度学习的高分辨率相位对比图像中亚细胞器的分割。
IF 2 4区 生物学
Cell structure and function Pub Date : 2024-08-30 Epub Date: 2024-07-31 DOI: 10.1247/csf.24036
Kentaro Shimasaki, Yuko Okemoto-Nakamura, Kyoko Saito, Masayoshi Fukasawa, Kaoru Katoh, Kentaro Hanada
{"title":"Deep learning-based segmentation of subcellular organelles in high-resolution phase-contrast images.","authors":"Kentaro Shimasaki, Yuko Okemoto-Nakamura, Kyoko Saito, Masayoshi Fukasawa, Kaoru Katoh, Kentaro Hanada","doi":"10.1247/csf.24036","DOIUrl":"10.1247/csf.24036","url":null,"abstract":"<p><p>Although quantitative analysis of biological images demands precise extraction of specific organelles or cells, it remains challenging in broad-field grayscale images, where traditional thresholding methods have been hampered due to complex image features. Nevertheless, rapidly growing artificial intelligence technology is overcoming obstacles. We previously reported the fine-tuned apodized phase-contrast microscopy system to capture high-resolution, label-free images of organelle dynamics in unstained living cells (Shimasaki, K. et al. (2024). Cell Struct. Funct., 49: 21-29). We here showed machine learning-based segmentation models for subcellular targeted objects in phase-contrast images using fluorescent markers as origins of ground truth masks. This method enables accurate segmentation of organelles in high-resolution phase-contrast images, providing a practical framework for studying cellular dynamics in unstained living cells.Key words: label-free imaging, organelle dynamics, apodized phase contrast, deep learning-based segmentation.</p>","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":" ","pages":"57-65"},"PeriodicalIF":2.0,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141859153","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Deciphering the sub-Golgi localization of glycosyltransferases via 3D super-resolution imaging. 通过三维超分辨率成像解密糖基转移酶的亚高尔基定位。
IF 2 4区 生物学
Cell structure and function Pub Date : 2024-08-27 Epub Date: 2024-08-22 DOI: 10.1247/csf.24008
Hirokazu Yagi, Seigo Tateo, Taiki Saito, Yusaku Ohta, Emiko Nishi, Saemi Obitsu, Tatsuya Suzuki, Supaphorn Seetaha, Charles Hellec, Akihiko Nakano, Takuro Tojima, Koichi Kato
{"title":"Deciphering the sub-Golgi localization of glycosyltransferases via 3D super-resolution imaging.","authors":"Hirokazu Yagi, Seigo Tateo, Taiki Saito, Yusaku Ohta, Emiko Nishi, Saemi Obitsu, Tatsuya Suzuki, Supaphorn Seetaha, Charles Hellec, Akihiko Nakano, Takuro Tojima, Koichi Kato","doi":"10.1247/csf.24008","DOIUrl":"10.1247/csf.24008","url":null,"abstract":"<p><p>The Golgi apparatus, a crucial organelle involved in protein processing, including glycosylation, exhibits complex sub-structures, i.e., cis-, medial, and trans-cisternae. This study investigated the distribution of glycosyltransferases within the Golgi apparatus of mammalian cells via 3D super-resolution imaging. Focusing on human glycosyltransferases involved in N-glycan modification, we found that even enzymes presumed to coexist in the same Golgi compartment exhibit nuanced variations in localization. By artificially making their N-terminal regions [composed of a cytoplasmic, transmembrane, and stem segment (CTS)] identical, it was possible to enhance the degree of their colocalization, suggesting the decisive role of this region in determining the sub-Golgi localization of enzymes. Ultimately, this study reveals the molecular codes within CTS regions as key determinants of glycosyltransferase localization, providing insights into precise control over the positioning of glycosyltransferases, and consequently, the interactions between glycosyltransferases and substrate glycoproteins as cargoes in the secretory pathway. This study advances our understanding of Golgi organization and opens avenues for programming the glycosylation of proteins for clinical applications.Key words: Golgi apparatus, glycosyltransferase, 3D super-resolution imaging, N-glycosylation.</p>","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":" ","pages":"47-55"},"PeriodicalIF":2.0,"publicationDate":"2024-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141579100","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Reconstitution of nuclear envelope subdomain formation on mitotic chromosomes in semi-intact cells 半贴壁细胞有丝分裂染色体上核膜亚域形成的重建
IF 1.5 4区 生物学
Cell structure and function Pub Date : 2024-06-04 DOI: 10.1247/csf.24003
Tomoko Funakoshi, Naoko Imamoto
{"title":"Reconstitution of nuclear envelope subdomain formation on mitotic chromosomes in semi-intact cells","authors":"Tomoko Funakoshi, Naoko Imamoto","doi":"10.1247/csf.24003","DOIUrl":"https://doi.org/10.1247/csf.24003","url":null,"abstract":"</p><p>In metazoans, the nuclear envelope (NE) disassembles during the prophase and reassembles around segregated chromatids during the telophase. The process of NE formation has been extensively studied using live-cell imaging. At the early step of NE reassembly in human cells, specific pattern-like localization of inner nuclear membrane (INM) proteins, connected to the nuclear pore complex (NPC), was observed in the so-called “core” region and “noncore” region on telophase chromosomes, which corresponded to the “pore-free” region and the “pore-rich” region, respectively, in the early G1 interphase nucleus. We refer to these phenomena as NE subdomain formation. To biochemically investigate this process, we aimed to develop an <i>in vitro</i> NE reconstitution system using digitonin-permeabilized semi-intact mitotic human cells coexpressing two INM proteins, emerin and lamin B receptor, which were labeled with fluorescent proteins. The targeting and accumulation of INM proteins to chromosomes before and after anaphase onset in semi-intact cells were observed using time-lapse imaging. Our <i>in vitro</i> NE reconstitution system recapitulated the formation of the NE subdomain, as in living cells, although chromosome segregation and cytokinesis were not observed. This <i>in vitro</i> NE reconstitution required the addition of a mitotic cytosolic fraction supplemented with a cyclin-dependent kinase inhibitor and energy sources. The cytoplasmic soluble factor(s) dependency of INM protein targeting differed among the segregation states of chromosomes. Furthermore, the NE reconstituted on segregated chromosomes exhibited active nucleocytoplasmic transport competency. These results indicate that the chromosome status changes after anaphase onset for recruiting NPC components.</p><p>","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"76 1","pages":""},"PeriodicalIF":1.5,"publicationDate":"2024-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141257791","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
相关产品
×
本文献相关产品
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信