{"title":"Adhesion-dependent Caveolin-1 Tyrosine-14 phosphorylation is regulated by FAK in response to changing matrix stiffness.","authors":"Natasha Buwa, Nivedhika Kannan, Shaunak Kanade, Nagaraj Balasubramanian","doi":"10.1002/1873-3468.14025","DOIUrl":"https://doi.org/10.1002/1873-3468.14025","url":null,"abstract":"<p><p>Integrin-mediated adhesion regulates cellular responses to changes in the mechanical and biochemical properties of the extracellular matrix. Cell-matrix adhesion regulates caveolar endocytosis, dependent on caveolin 1 (Cav1) Tyr14 phosphorylation (pY14Cav1), to control anchorage-dependent signaling. We find that cell-matrix adhesion regulates pY14Cav1 levels in mouse fibroblasts. Biochemical fractionation reveals endogenous pY14Cav1 to be present in caveolae and focal adhesions (FA). Adhesion does not affect caveolar pY14Cav1, supporting its regulation at FA, in which PF-228-mediated inhibition of focal adhesion kinase (FAK) disrupts. Cell adhesion on 2D polyacrylamide matrices of increasing stiffness stimulates Cav1 phosphorylation, which is comparable to the phosphorylation of FAK. Inhibition of FAK across varying stiffnesses shows it regulates pY14Cav1 more prominently at higher stiffness. Taken together, these studies reveal the presence of FAK-pY14Cav1 crosstalk at FA, which is regulated by cell-matrix adhesion.</p>","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":null,"pages":null},"PeriodicalIF":3.5,"publicationDate":"2021-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.14025","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38718389","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}
FEBS LettersPub Date : 2021-02-01Epub Date: 2020-12-08DOI: 10.1002/1873-3468.14008
Xiaoyan Cheng, Maolin Ge, Shouhai Zhu, Dan Li, Ruiheng Wang, Qiongyu Xu, Zhihong Chen, Shufeng Xie, Han Liu
{"title":"mTORC1-mediated amino acid signaling is critical for cell fate determination under transplant-induced stress.","authors":"Xiaoyan Cheng, Maolin Ge, Shouhai Zhu, Dan Li, Ruiheng Wang, Qiongyu Xu, Zhihong Chen, Shufeng Xie, Han Liu","doi":"10.1002/1873-3468.14008","DOIUrl":"https://doi.org/10.1002/1873-3468.14008","url":null,"abstract":"<p><p>Transplantation of in vitro-manipulated cells is widely used in hematology. While transplantation is well recognized to impose severe stress on transplanted cells, the nature of transplant-induced stress remains elusive. Here, we propose that the lack of amino acids in serum is the major cause of transplant-induced stress. Mechanistically, amino acid deficiency decreases protein synthesis and nutrient consummation. However, in cells with overactive AKT and ERK, mTORC1 is not inhibited and protein synthesis remains relatively high. This impaired signaling causes nutrient depletion, cell cycle block, and eventually autophagy and cell death, which can be inhibited by cycloheximide or mTORC1 inhibitors. Thus, mTORC1-mediated amino acid signaling is critical in cell fate determination under transplant-induced stress, and protein synthesis inhibition can improve transplantation efficiency.</p>","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":null,"pages":null},"PeriodicalIF":3.5,"publicationDate":"2021-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.14008","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38314118","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}
FEBS LettersPub Date : 2021-01-01Epub Date: 2020-11-13DOI: 10.1002/1873-3468.13978
Anusha Valpadashi, Sylvie Callegari, Andreas Linden, Piotr Neumann, Ralf Ficner, Henning Urlaub, Markus Deckers, Peter Rehling
{"title":"Defining the architecture of the human TIM22 complex by chemical crosslinking.","authors":"Anusha Valpadashi, Sylvie Callegari, Andreas Linden, Piotr Neumann, Ralf Ficner, Henning Urlaub, Markus Deckers, Peter Rehling","doi":"10.1002/1873-3468.13978","DOIUrl":"https://doi.org/10.1002/1873-3468.13978","url":null,"abstract":"<p><p>The majority of mitochondrial proteins are nuclear encoded and imported into mitochondria as precursor proteins via dedicated translocases. The translocase of the inner membrane 22 (TIM22) is a multisubunit molecular machine specialized for the translocation of hydrophobic, multi-transmembrane-spanning proteins with internal targeting signals into the inner mitochondrial membrane. Here, we undertook a crosslinking-mass spectrometry (XL-MS) approach to determine the molecular arrangement of subunits of the human TIM22 complex. Crosslinking of the isolated TIM22 complex using the BS3 crosslinker resulted in the broad generation of crosslinks across the majority of TIM22 components, including the small TIM chaperone complex. The crosslinking data uncovered several unexpected features, opening new avenues for a deeper investigation into the steps required for TIM22-mediated translocation in humans.</p>","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":null,"pages":null},"PeriodicalIF":3.5,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.13978","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38651007","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}
FEBS LettersPub Date : 2020-11-01Epub Date: 2020-09-10DOI: 10.1002/1873-3468.13915
Alexander Golubev, Bulat Fatkhullin, Iskander Khusainov, Lasse Jenner, Azat Gabdulkhakov, Shamil Validov, Gulnara Yusupova, Marat Yusupov, Konstantin Usachev
{"title":"Cryo-EM structure of the ribosome functional complex of the human pathogen Staphylococcus aureus at 3.2 Å resolution.","authors":"Alexander Golubev, Bulat Fatkhullin, Iskander Khusainov, Lasse Jenner, Azat Gabdulkhakov, Shamil Validov, Gulnara Yusupova, Marat Yusupov, Konstantin Usachev","doi":"10.1002/1873-3468.13915","DOIUrl":"https://doi.org/10.1002/1873-3468.13915","url":null,"abstract":"<p><p>Staphylococcus aureus is a bacterial pathogen and one of the leading causes of healthcare-acquired infections in the world. The growing antibiotic resistance of S. aureus obliges us to search for new drugs and treatments. As the majority of antibiotics target the ribosome, knowledge of its detailed structure is crucial for drug development. Here, we report the cryo-EM reconstruction at 3.2 Å resolution of the S. aureus ribosome with P-site tRNA, messenger RNA, and 10 RNA modification sites previously not assigned or visualized. The resulting model is the most precise and complete high-resolution structure to date of the S. aureus 70S ribosome with functional ligands.</p>","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":null,"pages":null},"PeriodicalIF":3.5,"publicationDate":"2020-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.13915","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38315747","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}
FEBS LettersPub Date : 2020-11-01Epub Date: 2020-09-20DOI: 10.1002/1873-3468.13924
Tadashi Nakaya
{"title":"Dissection of FUS domains involved in regulation of SnRNP70 gene expression.","authors":"Tadashi Nakaya","doi":"10.1002/1873-3468.13924","DOIUrl":"https://doi.org/10.1002/1873-3468.13924","url":null,"abstract":"<p><p>FUS is one of the causative factors of amyotrophic lateral sclerosis. Loss and/or gain of its physiological functions has been believed to be linked to the pathogenesis of this condition. However, its functions remain incompletely understood. This study dissected the domains of FUS regulating the expression of SnRNP70, which functions in mRNA splicing. Biochemical analysis revealed that all FUS domains except for RGG1 contribute to determining Snrnp70 transcript abundance and thus its protein abundance. RNA-Seq analysis using the Gly-rich domain-deleted mutant coupled with snRNP70 knockdown revealed that FUS has a potential to regulate gene expression in both snRNP70-dependent and snRNP70-independent manners through the Gly-rich domain. These results provide insight into molecular details of the regulation of gene expression by FUS.</p>","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":null,"pages":null},"PeriodicalIF":3.5,"publicationDate":"2020-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.13924","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38370244","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}
FEBS LettersPub Date : 2020-11-01Epub Date: 2020-09-04DOI: 10.1002/1873-3468.13913
Ekaterina A Vasilenko, Ekaterina N Gorshkova, Irina V Astrakhantseva, Marina S Drutskaya, Sergei V Tillib, Sergei A Nedospasov, Vladislav V Mokhonov
{"title":"The structure of myeloid cell-specific TNF inhibitors affects their biological properties.","authors":"Ekaterina A Vasilenko, Ekaterina N Gorshkova, Irina V Astrakhantseva, Marina S Drutskaya, Sergei V Tillib, Sergei A Nedospasov, Vladislav V Mokhonov","doi":"10.1002/1873-3468.13913","DOIUrl":"https://doi.org/10.1002/1873-3468.13913","url":null,"abstract":"<p><p>Spatial organization and conformational changes of antibodies may significantly affect their biological functions. We assessed the effect of mutual organization of the two V<sub>H</sub> H domains within bispecific antibodies recognizing human TNF and the surface molecules of murine myeloid cells (F4/80 or CD11b) on TNF retention and inhibition. TNF-neutralizing properties in vitro and in vivo of MYSTI-2 and MYSTI-3 antibodies were compared with new variants with interchanged V<sub>H</sub> H domains and different linker sequences. The most effective structure of MYSTI-2 and MYSTI-3 proteins required the Ser/Gly-containing 'superflexible' linker. The orientation of the modules was crucial for the activity of the proteins, but not for MYSTI-3 with the Pro/Gln-containing 'semi-rigid' linker. Our results may contribute toward the development of more effective drug prototypes.</p>","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":null,"pages":null},"PeriodicalIF":3.5,"publicationDate":"2020-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.13913","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38422094","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}
FEBS LettersPub Date : 2020-11-01Epub Date: 2020-09-13DOI: 10.1002/1873-3468.13920
Krithika Rajagopalan, Jonathan Dworkin
{"title":"Escherichia coli YegI is a novel Ser/Thr kinase lacking conserved motifs that localizes to the inner membrane.","authors":"Krithika Rajagopalan, Jonathan Dworkin","doi":"10.1002/1873-3468.13920","DOIUrl":"https://doi.org/10.1002/1873-3468.13920","url":null,"abstract":"<p><p>In bacteria, signaling phosphorylation is thought to occur primarily on His and Asp residues. However, phosphoproteomic surveys in phylogenetically diverse bacteria over the past decade have identified numerous proteins that are phosphorylated on Ser and/or Thr residues. Consistently, genes encoding Ser/Thr kinases are present in many bacterial genomes such as in the Escherichia coli genome, which encodes at least three Ser/Thr kinases. Here, we identify a previously uncharacterized ORF, yegI, and demonstrate that it encodes a novel Ser/Thr kinase. YegI lacks several conserved motifs including residues important for Mg<sup>2+</sup> binding seen in other bacterial Ser/Thr kinases, suggesting that the consensus may be too stringent. We further find that YegI is a two-pass membrane protein with both N- and C termini located intracellularly.</p>","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":null,"pages":null},"PeriodicalIF":3.5,"publicationDate":"2020-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.13920","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38347945","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}
FEBS LettersPub Date : 2020-11-01Epub Date: 2020-09-18DOI: 10.1002/1873-3468.13923
Jing Jin, Ze-Hua Guo, Quan Hao, Mee-Len Chye
{"title":"Crystal structure of the rice acyl-CoA-binding protein OsACBP2 in complex with C18:3-CoA reveals a novel pattern of binding to acyl-CoA esters.","authors":"Jing Jin, Ze-Hua Guo, Quan Hao, Mee-Len Chye","doi":"10.1002/1873-3468.13923","DOIUrl":"https://doi.org/10.1002/1873-3468.13923","url":null,"abstract":"Acyl‐CoA‐binding proteins (ACBPs) are a family of proteins that bind acyl‐CoA esters at a conserved acyl‐CoA‐binding domain. ACBPs maintain intracellular acyl‐CoA pools to regulate lipid metabolism. Here, we report on the structure of rice OsACBP2 in complex with C18:3‐CoA ester. The residues Y33, K34 and K56 of OsACBP2 play a crucial role in binding the CoA group, while residues N23, L27, K52 and Y55 in one molecule of OsACBP2 cooperate with L27, L28, A59 and A62 from another anchoring the fatty acyl group. Multiangle light scattering assays indicate that OsACBP2 binds C18:3‐CoA as a monomer. The first complex structure of a plant ACBP binding with C18:3‐CoA is therefore presented, providing a novel model for the interaction between an acyl‐CoA ester and the acyl‐CoA‐binding domain(s).","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":null,"pages":null},"PeriodicalIF":3.5,"publicationDate":"2020-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.13923","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38346317","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}
FEBS LettersPub Date : 2020-07-01Epub Date: 2020-05-31DOI: 10.1002/1873-3468.13808
Petra Diestelkoetter-Bachert, Rainer Beck, Inge Reckmann, Andrea Hellwig, Ana Garcia-Saez, Monika Zelman-Hopf, Anton Hanke, Ariane Nunes Alves, Rebecca C Wade, Matthias P Mayer, Felix Wieland
{"title":"Structural characterization of an Arf dimer interface: molecular mechanism of Arf-dependent membrane scission.","authors":"Petra Diestelkoetter-Bachert, Rainer Beck, Inge Reckmann, Andrea Hellwig, Ana Garcia-Saez, Monika Zelman-Hopf, Anton Hanke, Ariane Nunes Alves, Rebecca C Wade, Matthias P Mayer, Felix Wieland","doi":"10.1002/1873-3468.13808","DOIUrl":"https://doi.org/10.1002/1873-3468.13808","url":null,"abstract":"<p><p>Dimerization of the small GTPase Arf is prerequisite for the scission of COPI-coated transport vesicles. Here, we quantify the monomer/dimer equilibrium of Arf within the membrane and show that after membrane scission, Arf dimers are restricted to donor membranes. By hydrogen exchange mass spectrometry, we define the interface of activated dimeric Arf within its switch II region. Single amino acid exchanges in this region reduce the propensity of Arf to dimerize. We suggest a mechanism of membrane scission by which the dimeric form of Arf is segregated to the donor membrane. Our data are consistent with the previously reported absence of dimerized Arf in COPI vesicles and could explain the presence of one single scar-like noncoated region in each COPI vesicle.</p>","PeriodicalId":50454,"journal":{"name":"FEBS Letters","volume":null,"pages":null},"PeriodicalIF":3.5,"publicationDate":"2020-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1873-3468.13808","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37924849","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}