European journal of cell biology最新文献

{"title":"The solute carrier SLC7A1 may act as a protein transporter at the blood-brain barrier","authors":"Magdalena Kurtyka ,&nbsp;Frank Wessely ,&nbsp;Sarah Bau ,&nbsp;Eseoghene Ifie ,&nbsp;Liqun He ,&nbsp;Nienke M. de Wit ,&nbsp;Alberte Bay Villekjær Pedersen ,&nbsp;Maximilian Keller ,&nbsp;Caleb Webber ,&nbsp;Helga E. de Vries ,&nbsp;Olaf Ansorge ,&nbsp;Christer Betsholtz ,&nbsp;Marijke De Bock ,&nbsp;Catarina Chaves ,&nbsp;Birger Brodin ,&nbsp;Morten S. Nielsen ,&nbsp;Winfried Neuhaus ,&nbsp;Robert D. Bell ,&nbsp;Tamás Letoha ,&nbsp;Axel H. Meyer ,&nbsp;Claus U. Pietrzik","doi":"10.1016/j.ejcb.2024.151406","DOIUrl":"10.1016/j.ejcb.2024.151406","url":null,"abstract":"<div><p>Despite extensive research, targeted delivery of substances to the brain still poses a great challenge due to the selectivity of the blood-brain barrier (BBB). Most molecules require either carrier- or receptor-mediated transport systems to reach the central nervous system (CNS). These transport systems form attractive routes for the delivery of therapeutics into the CNS, yet the number of known brain endothelium-enriched receptors allowing the transport of large molecules into the brain is scarce. Therefore, to identify novel BBB targets, we combined transcriptomic analysis of human and murine brain endothelium and performed a complex screening of BBB-enriched genes according to established selection criteria. As a result, we propose the high-affinity cationic amino acid transporter 1 (SLC7A1) as a novel candidate for transport of large molecules across the BBB. Using RNA sequencing and in situ hybridization assays, we demonstrated elevated <em>SLC7A1</em> gene expression in both human and mouse brain endothelium. Moreover, we confirmed SLC7A1 protein expression in brain vasculature of both young and aged mice. To assess the potential of SLC7A1 as a transporter for larger proteins, we performed internalization and transcytosis studies using a radiolabelled or fluorophore-labelled anti-SLC7A1 antibody. Our results showed that SLC7A1 internalised a SLC7A1-specific antibody in human colorectal carcinoma (HCT116) cells. Moreover, transcytosis studies in both immortalised human brain endothelial (hCMEC/D3) cells and primary mouse brain endothelial cells clearly demonstrated that SLC7A1 effectively transported the SLC7A1-specific antibody from luminal to abluminal side. Therefore, here in this study, we present for the first time the SLC7A1 as a novel candidate for transport of larger molecules across the BBB.</p></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 2","pages":"Article 151406"},"PeriodicalIF":6.6,"publicationDate":"2024-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0171933524000232/pdfft?md5=f35f1d0d69bba4a3f28797ca29dc4ad1&pid=1-s2.0-S0171933524000232-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140277743","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Arp2/3 complex- and formin-mediated actin cytoskeleton networks facilitate actin binding protein sorting in fission yeast","authors":"Kaitlin E. Homa ,&nbsp;Glen M. Hocky ,&nbsp;Cristian Suarez ,&nbsp;David R. Kovar","doi":"10.1016/j.ejcb.2024.151404","DOIUrl":"https://doi.org/10.1016/j.ejcb.2024.151404","url":null,"abstract":"<div><p>While it is well-established that F-actin networks with specific organizations and dynamics are tightly regulated by distinct sets of associated actin-binding proteins (ABPs), how ABPs self-sort to particular F-actin networks remains largely unclear. We report that actin assembly factors Arp2/3 complex and formin Cdc12 tune the association of ABPs fimbrin Fim1 and tropomyosin Cdc8 to different F-actin networks in fission yeast. Genetic and pharmacological disruption of F-actin networks revealed that Fim1 is preferentially directed to Arp2/3-complex mediated actin patches, whereas Cdc8 is preferentially targeted to formin Cdc12-mediated filaments in the contractile ring. To investigate the role of Arp2/3 complex- and formin Cdc12-mediated actin assembly, we used four-color TIRF microscopy to observe the in vitro reconstitution of ABP sorting with purified proteins. Fim1 or Cdc8 alone bind similarly well to filaments assembled by either assembly factor. However, in ‘competition’ reactions containing both actin assembly factors and both ABPs, ∼2.0-fold more Fim1 and ∼3.5-fold more Cdc8 accumulates on Arp2/3 complex branch points and formin Cdc12-assembled actin filaments, respectively. These findings indicate that F-actin assembly factors Arp2/3 complex and formin Cdc12 help facilitate the recruitment of specific ABPs, thereby tuning ABP sorting and subsequently establishing the identity of F-actin networks in fission yeast.</p></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 2","pages":"Article 151404"},"PeriodicalIF":6.6,"publicationDate":"2024-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0171933524000219/pdfft?md5=8304ed8acd8b10fb628417601482cee3&pid=1-s2.0-S0171933524000219-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140139026","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The small yeast GTPase Rho5 requires specific mitochondrial outer membrane proteins for translocation under oxidative stress and interacts with the VDAC Por1","authors":"Linnet Bischof,&nbsp;Franziska Schweitzer,&nbsp;Hans-Peter Schmitz,&nbsp;Jürgen J. Heinisch","doi":"10.1016/j.ejcb.2024.151405","DOIUrl":"https://doi.org/10.1016/j.ejcb.2024.151405","url":null,"abstract":"<div><p>Yeast Rho5 is a small GTPase which mediates the response to nutrient and oxidative stress, and triggers mitophagy and apoptosis. We here studied the rapid translocation of a GFP-tagged Rho5 to mitochondria under such stress conditions by live-cell fluorescence microscopy in the background of strains lacking different mitochondrial outer membrane proteins (MOMP). Fun14, Msp1 and Alo1 were found to be required for efficient recruitment of the GTPase, whereas translocation of Dck1 and Lmo1, the subunits of its dimeric GDP/GTP exchange factor (GEF), remained unaffected. An influence of the voltage-dependent anion channel (VDAC) Por1 on the association of GFP-Rho5 with mitochondria under oxidative stress conditions appeared to be strain-dependent. However, epistasis analyses and bimolecular fluorescence complementation (BiFC) studies indicate a genetic and physical interaction. All four strains lacking a single MOMP were investigated for their effect on mitophagy.</p></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 2","pages":"Article 151405"},"PeriodicalIF":6.6,"publicationDate":"2024-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0171933524000220/pdfft?md5=2d55ac6b32853bfa5e71c7b6ab106a18&pid=1-s2.0-S0171933524000220-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140163207","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Reconstitution of cytolinker-mediated crosstalk between actin and vimentin","authors":"Irene Istúriz Petitjean ,&nbsp;Quang D. Tran ,&nbsp;Angeliki Goutou ,&nbsp;Zima Kabir ,&nbsp;Gerhard Wiche ,&nbsp;Cécile Leduc ,&nbsp;Gijsje H. Koenderink","doi":"10.1016/j.ejcb.2024.151403","DOIUrl":"10.1016/j.ejcb.2024.151403","url":null,"abstract":"<div><p>Cell shape and motility are determined by the cytoskeleton, an interpenetrating network of actin filaments, microtubules, and intermediate filaments. The biophysical properties of each filament type individually have been studied extensively by cell-free reconstitution. By contrast, the interactions between the three cytoskeletal networks are relatively unexplored. They are coupled via crosslinkers of the plakin family such as plectin. These are challenging proteins for reconstitution because of their giant size and multidomain structure. Here we engineer a recombinant actin-vimentin crosslinker protein called ‘ACTIF’ that provides a minimal model system for plectin, recapitulating its modular design with actin-binding and intermediate filament-binding domains separated by a coiled-coil linker for dimerisation. We show by fluorescence and electron microscopy that ACTIF has a high binding affinity for vimentin and actin and creates mixed actin-vimentin bundles. Rheology measurements show that ACTIF-mediated crosslinking strongly stiffens actin-vimentin composites. Finally, we demonstrate the modularity of this approach by creating an ACTIF variant with the intermediate filament binding domain of Adenomatous Polyposis Coli. Our protein engineering approach provides a new cell-free system for the biophysical characterization of intermediate filament-binding crosslinkers and for understanding the mechanical synergy between actin and vimentin in mesenchymal cells.</p></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 2","pages":"Article 151403"},"PeriodicalIF":6.6,"publicationDate":"2024-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0171933524000207/pdfft?md5=451d043d0bc0cda6e2659ddfd35dccd1&pid=1-s2.0-S0171933524000207-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140128663","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Actin-membrane linkers: Insights from synthetic reconstituted systems","authors":"Feng-Ching Tsai ,&nbsp;Gwendal Guérin ,&nbsp;Julien Pernier ,&nbsp;Patricia Bassereau","doi":"10.1016/j.ejcb.2024.151402","DOIUrl":"10.1016/j.ejcb.2024.151402","url":null,"abstract":"<div><p>At the cell surface, the actin cytoskeleton and the plasma membrane interact reciprocally in a variety of processes related to the remodeling of the cell surface. The actin cytoskeleton has been known to modulate membrane organization and reshape the membrane. To this end, actin-membrane linking molecules play a major role in regulating actin assembly and spatially direct the interaction between the actin cytoskeleton and the membrane. While studies in cells have provided a wealth of knowledge on the molecular composition and interactions of the actin-membrane interface, the complex molecular interactions make it challenging to elucidate the precise actions of the actin-membrane linkers at the interface. Synthetic reconstituted systems, consisting of model membranes and purified proteins, have been a powerful approach to elucidate how actin-membrane linkers direct actin assembly to drive membrane shape changes. In this review, we will focus only on several actin-membrane linkers that have been studied by using reconstitution systems. We will discuss the design principles of these reconstitution systems and how they have contributed to the understanding of the cellular functions of actin-membrane linkers. Finally, we will provide a perspective on future research directions in understanding the intricate actin-membrane interaction.</p></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 2","pages":"Article 151402"},"PeriodicalIF":6.6,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0171933524000190/pdfft?md5=57b937c8a7a0cf9a66db72e1c542da38&pid=1-s2.0-S0171933524000190-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140047310","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cross-regulation of Listeria monocytogenes and the host ubiquitin system in listeriosis","authors":"Yuan Zhuang ,&nbsp;Johanna B. Fischer ,&nbsp;Gopala Nishanth ,&nbsp;Dirk Schlüter","doi":"10.1016/j.ejcb.2024.151401","DOIUrl":"10.1016/j.ejcb.2024.151401","url":null,"abstract":"<div><p>The facultative intracellular bacterium <em>Listeria</em> (<em>L</em>.) <em>monocytogenes</em> may cause severe diseases in humans and animals. The control of listeriosis/<em>L. monocytogenes</em> requires the concerted action of cells of the innate and adaptive immune systems. In this regard, cell-intrinsic immunity of infected cells, activated by the immune responses, is crucial for the control and elimination intracellular <em>L. monocytogenes</em>. Both the immune response against <em>L. monocytogenes</em> and cell intrinsic pathogen control are critically regulated by post-translational modifications exerted by the host ubiquitin system and ubiquitin-like modifiers (Ubls). In this review, we discuss our current understanding of the role of the ubiquitin system and Ubls in listeriosis, as well as future directions of research.</p></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 2","pages":"Article 151401"},"PeriodicalIF":6.6,"publicationDate":"2024-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0171933524000189/pdfft?md5=2e8603c47657a372fbad6143dc985edf&pid=1-s2.0-S0171933524000189-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140007916","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"NANOS1 restricts oral cancer cell motility and TGF-ß signaling","authors":"Julia Rosemann ,&nbsp;Jonas Pyko ,&nbsp;Roland Jacob ,&nbsp;Jana Macho ,&nbsp;Matthias Kappler ,&nbsp;Alexander W. Eckert ,&nbsp;Monika Haemmerle ,&nbsp;Tony Gutschner","doi":"10.1016/j.ejcb.2024.151400","DOIUrl":"https://doi.org/10.1016/j.ejcb.2024.151400","url":null,"abstract":"<div><p>Oral squamous cell carcinoma (OSCC) is the most frequent type of cancer of the head and neck area accounting for approx. 377,000 new cancer cases every year. The epithelial-to-mesenchymal transition (EMT) program plays an important role in OSCC progression and metastasis therefore contributing to a poor prognosis in patients with advanced disease. Transforming growth factor beta (TGF-ß) is a powerful inducer of EMT thereby increasing cancer cell aggressiveness. Here, we aimed at identifying RNA-binding proteins (RBPs) that affect TGF-ß-induced EMT. To this end we treated oral cancer cells with TGF-ß and identified a total of 643 significantly deregulated protein-coding genes in response to TGF-ß. Of note, 19 genes encoded RBPs with NANOS1 being the most downregulated RBP. Subsequent cellular studies demonstrated a strong inhibitory effect of NANOS1 on migration and invasion of SAS oral cancer cells. Further mechanistic studies revealed an interaction of NANOS1 with the TGF-ß receptor 1 (TGFBR1) mRNA, leading to increased decay of this transcript and a reduced TGFBR1 protein expression, thereby preventing downstream TGF-ß/SMAD signaling. In summary, we identified NANOS1 as negative regulator of TGF-ß signaling in oral cancer cells.</p></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 2","pages":"Article 151400"},"PeriodicalIF":6.6,"publicationDate":"2024-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0171933524000177/pdfft?md5=167b7a8af9a06c0c2500eddda780aba1&pid=1-s2.0-S0171933524000177-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139942605","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Immortalised murine R349P desmin knock-in myotubes exhibit a reduced proton leak and decreased ADP/ATP translocase levels in purified mitochondria","authors":"Carolin Berwanger ,&nbsp;Dominic Terres ,&nbsp;Dominik Pesta ,&nbsp;Britta Eggers ,&nbsp;Katrin Marcus ,&nbsp;Ilka Wittig ,&nbsp;Rudolf J. Wiesner ,&nbsp;Rolf Schröder ,&nbsp;Christoph S. Clemen","doi":"10.1016/j.ejcb.2024.151399","DOIUrl":"10.1016/j.ejcb.2024.151399","url":null,"abstract":"<div><p>Desmin gene mutations cause myopathies and cardiomyopathies. Our previously characterised R349P desminopathy mice, which carry the ortholog of the common human desmin mutation R350P, showed marked alterations in mitochondrial morphology and function in muscle tissue. By isolating skeletal muscle myoblasts from offspring of R349P desminopathy and p53 knock-out mice, we established an immortalised cellular disease model. Heterozygous and homozygous R349P desmin knock-in and wild-type myoblasts could be well differentiated into multinucleated spontaneously contracting myotubes. The desminopathy myoblasts showed the characteristic disruption of the desmin cytoskeleton and desmin protein aggregation, and the desminopathy myotubes showed the characteristic myofibrillar irregularities. Long-term electrical pulse stimulation promoted myotube differentiation and markedly increased their spontaneous contraction rate. In both heterozygous and homozygous R349P desminopathy myotubes, this treatment restored a regular myofibrillar cross-striation pattern as seen in wild-type myotubes. High-resolution respirometry of mitochondria purified from myotubes by density gradient ultracentrifugation revealed normal oxidative phosphorylation capacity, but a significantly reduced proton leak in mitochondria from the homozygous R349P desmin knock-in cells. Consistent with a reduced proton flux across the inner mitochondrial membrane, our quantitative proteomic analysis of the purified mitochondria revealed significantly reduced levels of ADP/ATP translocases in the homozygous R349P desmin knock-in genotype. As this alteration was also detected in the soleus muscle of R349P desminopathy mice, which, in contrast to the mitochondria purified from cultured cells, showed a variety of other dysregulated mitochondrial proteins, we consider this finding to be an early step in the pathogenesis of secondary mitochondriopathy in desminopathy.</p></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 2","pages":"Article 151399"},"PeriodicalIF":6.6,"publicationDate":"2024-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0171933524000165/pdfft?md5=d463031335b9d8d2f66af0b6b1840115&pid=1-s2.0-S0171933524000165-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139920196","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular mechanisms of naringenin modulation of mitochondrial permeability transition acting on F1FO-ATPase and counteracting saline load-induced injury in SHRSP cerebral endothelial cells","authors":"Salvatore Nesci ,&nbsp;Cristina Algieri ,&nbsp;Matteo Antonio Tallarida ,&nbsp;Rosita Stanzione ,&nbsp;Saverio Marchi ,&nbsp;Donatella Pietrangelo ,&nbsp;Fabiana Trombetti ,&nbsp;Luca D’Ambrosio ,&nbsp;Maurizio Forte ,&nbsp;Maria Cotugno ,&nbsp;Ilaria Nunzi ,&nbsp;Rachele Bigi ,&nbsp;Loredana Maiuolo ,&nbsp;Antonio De Nino ,&nbsp;Paolo Pinton ,&nbsp;Giovanni Romeo ,&nbsp;Speranza Rubattu","doi":"10.1016/j.ejcb.2024.151398","DOIUrl":"https://doi.org/10.1016/j.ejcb.2024.151398","url":null,"abstract":"<div><p>Naringenin (NRG) was characterized for its ability to counteract mitochondrial dysfunction which is linked to cardiovascular diseases. The F₁F_O-ATPase can act as a molecular target of NRG. The interaction of NRG with this enzyme can avoid the energy transmission mechanism of ATP hydrolysis, especially in the presence of Ca²⁺ cation used as cofactor. Indeed, NRG was a selective inhibitor of the hydrophilic F₁ domain displaying a binding site overlapped with quercetin in the inside surface of an annulus made by the three α and the three β subunits arranged alternatively in a hexamer. The kinetic constant of inhibition suggested that NRG preferred the enzyme activated by Ca²⁺ rather than the F₁F_O-ATPase activated by the natural cofactor Mg²⁺. From the inhibition type mechanism of NRG stemmed the possibility to speculate that NRG can prevent the activation of F₁F_O-ATPase by Ca²⁺. The event correlated to the protective role in the mitochondrial permeability transition pore opening by NRG as well as to the reduction of ROS production probably linked to the NRG chemical structure with antioxidant action. Moreover, in primary cerebral endothelial cells (ECs) obtained from stroke prone spontaneously hypertensive rats NRG had a protective effect on salt-induced injury by restoring cell viability and endothelial cell tube formation while also rescuing complex I activity.</p></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 2","pages":"Article 151398"},"PeriodicalIF":6.6,"publicationDate":"2024-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0171933524000153/pdfft?md5=44a7a4ec51bf85a8291fe739ca43f7a5&pid=1-s2.0-S0171933524000153-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139749326","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Deletion of exons 2 and 3 from Actb and cell immortalization lead to widespread, β-actin independent alterations in gene expression associated with cell cycle control","authors":"Lauren J. Sundby ,&nbsp;William M. Southern ,&nbsp;Jiao Sun ,&nbsp;Xiaobai Patrinostro ,&nbsp;Wei Zhang ,&nbsp;Jeongsik Yong ,&nbsp;James M. Ervasti","doi":"10.1016/j.ejcb.2024.151397","DOIUrl":"10.1016/j.ejcb.2024.151397","url":null,"abstract":"<div><p>The cytoplasmic actin proteins, β- and γ-actin, are 99% identical but thought to perform non-redundant functions. The nucleotide coding regions of cytoplasmic actin genes, <em>Actb</em> and <em>Actg1</em>, are 89% identical. Knockout (KO) of <em>Actb</em> by Cre-mediated deletion of first coding exons 2 and 3 in mice is embryonic lethal and fibroblasts derived from KO embryos (MEFs) fail to proliferate. In contrast, <em>Actg1</em> KO MEFs display with a much milder defect in cell proliferation and <em>Actg1</em> KO mice are viable, but present with increased perinatal lethality. Recent studies have identified important protein-independent functions for both <em>Actb</em> and <em>Actg1</em> and demonstrate that deletions within the <em>Actb</em> nucleotide sequence, and not loss of the β-actin protein, cause the most severe phenotypes in KO mice and cells. Here, we use a multi-omics approach to better understand what drives the phenotypes of <em>Actb</em> KO MEFs. RNA-sequencing and mass spectrometry reveal largescale changes to the transcriptome, proteome, and phosphoproteome in cells lacking <em>Actb</em> but not those only lacking β-actin protein. Pathway analysis of genes and proteins differentially expressed upon <em>Actb</em> KO suggest widespread dysregulation of genes involved in the cell cycle that may explain the severe defect in proliferation.</p></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 2","pages":"Article 151397"},"PeriodicalIF":6.6,"publicationDate":"2024-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0171933524000141/pdfft?md5=e1f8394c38e517d78b6126ca98964fca&pid=1-s2.0-S0171933524000141-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139827702","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}