المؤلفون: Ocasio, Cory A., Baggelaar, Marc P., Sipthorp, James, Losada de la Lastra, Ana, Tavares, Manuel, Volarić, Jana, Soudy, Christelle, Storck, Elisabeth M., Houghton, Jack W., Palma-Duran, Susana A., MacRae, James I., Tomić, Goran, Carr, Lotte, Downward, Julian, Eggert, Ulrike S., Tate, Edward W.

المساهمون: Cancer Research UK, Wellcome Trust, European Research Council Advanced Grant RASImmune, RCUK | Engineering and Physical Sciences Research Council

المصدر: Nature Biotechnology ; ISSN 1087-0156 1546-1696

مصطلحات موضوعية: Biomedical Engineering, Molecular Medicine, Applied Microbiology and Biotechnology, Bioengineering, Biotechnology

الوصف: The 23 human zinc finger Asp-His-His-Cys motif-containing (ZDHHC) S -acyltransferases catalyze long-chain S -acylation at cysteine residues across an extensive network of hundreds of proteins important for normal physiology or dysregulated in disease. Here we present a technology to directly map the protein substrates of a specific ZDHHC at the whole-proteome level, in intact cells. Structure-guided engineering of paired ZDHHC ‘hole’ mutants and ‘bumped’ chemically tagged fatty acid probes enabled probe transfer to specific protein substrates with excellent selectivity over wild-type ZDHHCs. Chemical–genetic systems were exemplified for five human ZDHHCs (3, 7, 11, 15 and 20) and applied to generate de novo ZDHHC substrate profiles, identifying >300 substrates and S -acylation sites for new functionally diverse proteins across multiple cell lines. We expect that this platform will elucidate S -acylation biology for a wide range of models and organisms.

الإتاحة: https://doi.org/10.1038/s41587-023-02030-0Test
https://www.nature.com/articles/s41587-023-02030-0.pdfTest
https://www.nature.com/articles/s41587-023-02030-0Test

المؤلفون: Méndez-Lucas, Andrés, Lin, Wei, Driscoll, Paul C, Legrave, Nathalie, Novellasdemunt, Laura, Xie, Chencheng, Charles, Mark, Wilson, Zena, Jones, Neil P, Rayport, Stephen, Rodríguez-Justo, Manuel, Li, Vivian, MacRae, James I, Hay, Nissim, Chen, Xin, Yuneva, Mariia

المصدر: Nature Metabolism. 2(4)

مصطلحات موضوعية: Medical Biochemistry and Metabolomics, Medical Physiology, Biomedical and Clinical Sciences, Nutrition and Dietetics, Nutrition, 5.1 Pharmaceuticals, Development of treatments and therapeutic interventions, 2.1 Biological and endogenous factors, Aetiology, Cancer, Animals, Cell Proliferation, Glucose, Glutaminase, Glutamine, Humans, Liver Neoplasms, Mice, Proto-Oncogene Proteins c-myc, Medical biochemistry and metabolomics, Medical physiology, Nutrition and dietetics

الوصف: Plasticity of cancer metabolism can be a major obstacle to efficient targeting of tumour-specific metabolic vulnerabilities. Here, we identify the compensatory mechanisms following the inhibition of major pathways of central carbon metabolism in c-MYC-induced liver tumours. We find that, while inhibition of both glutaminase isoforms (Gls1 and Gls2) in tumours considerably delays tumourigenesis, glutamine catabolism continues, owing to the action of amidotransferases. Synergistic inhibition of both glutaminases and compensatory amidotransferases is required to block glutamine catabolism and proliferation of mouse and human tumour cells in vitro and in vivo. Gls1 deletion is also compensated for by glycolysis. Thus, co-inhibition of Gls1 and hexokinase 2 significantly affects Krebs cycle activity and tumour formation. Finally, the inhibition of biosynthesis of either serine (Psat1-KO) or fatty acid (Fasn-KO) is compensated for by uptake of circulating nutrients, and dietary restriction of both serine and glycine or fatty acids synergistically suppresses tumourigenesis. These results highlight the high flexibility of tumour metabolism and demonstrate that either pharmacological or dietary targeting of metabolic compensatory mechanisms can improve therapeutic outcomes.

وصف الملف: application/pdf

الوصول الحر: https://escholarship.org/uc/item/7mf8j1hkTest

المؤلفون: Giampazolias, Evangelos, da Costa, Mariana Pereira, Lam, Khiem C, Lim, Kok Haw Jonathan, Cardoso, Ana, Piot, Cécile, Chakravarty, Probir, Blasche, Sonja, Patel, Swara, Biram, Adi, Castro-Dopico, Tomas, Buck, Michael D, Rodrigues, Richard R, Poulsen, Gry Juul, Palma-Duran, Susana A, Rogers, Neil C, Koufaki, Maria A, Minutti, Carlos M, Wang, Pengbo, Vdovin, Alexander, Frederico, Bruno, Childs, Eleanor, Lee, Sonia, Simpson, Ben, Iseppon, Andrea, Omenetti, Sara, Kelly, Gavin, Goldstone, Robert, Nye, Emma, Suárez-Bonnet, Alejandro, Priestnall, Simon L, MacRae, James I, Zelenay, Santiago, Patil, Kiran Raosaheb, Litchfield, Kevin, Lee, James C, Jess, Tine, Goldszmid, Romina S, Reis e Sousa, Caetano

مصطلحات موضوعية: Immunology, FOS Clinical medicine, Tumour Biology

الوصف: A role for vitamin D in immune modulation and in cancer has been suggested. In this work, we report that mice with increased availability of vitamin D display greater immune-dependent resistance to transplantable cancers and augmented responses to checkpoint blockade immunotherapies. Similarly, in humans, vitamin D-induced genes correlate with improved responses to immune checkpoint inhibitor treatment as well as with immunity to cancer and increased overall survival. In mice, resistance is attributable to the activity of vitamin D on intestinal epithelial cells, which alters microbiome composition in favor of Bacteroides fragilis, which positively regulates cancer immunity. Our findings indicate a previously unappreciated connection between vitamin D, microbial commensal communities, and immune responses to cancer. Collectively, they highlight vitamin D levels as a potential determinant of cancer immunity and immunotherapy success. ...

الإتاحة: https://doi.org/10.25418/crick.25772151Test
https://crick.figshare.com/articles/journal_contribution/Vitamin_D_regulates_microbiome-dependent_cancer_immunity_/25772151Test

المؤلفون: Amodeo, Valeria, Davies, Timothy, Martinez-Segura, Amalia, Clements, Melanie P, Ragdale, Holly Simpson, Bailey, Andrew, Dos Santos, Mariana Silva, MacRae, James I, Mokochinski, Joao, Kramer, Holger, Garcia-Diaz, Claudia, Gould, Alex P, Marguerat, Samuel, Parrinello, Simona

المصدر: Developmental Cell (2023) (In press).

مصطلحات موضوعية: fatty acid oxidation, metabolism, neural stem cells, p53, quiescence, tumour initiation

الوصف: Glioblastoma is thought to originate from neural stem cells (NSCs) of the subventricular zone that acquire genetic alterations. In the adult brain, NSCs are largely quiescent, suggesting that deregulation of quiescence maintenance may be a prerequisite for tumor initiation. Although inactivation of the tumor suppressor p53 is a frequent event in gliomagenesis, whether or how it affects quiescent NSCs (qNSCs) remains unclear. Here, we show that p53 maintains quiescence by inducing fatty-acid oxidation (FAO) and that acute p53 deletion in qNSCs results in their premature activation to a proliferative state. Mechanistically, this occurs through direct transcriptional induction of PPARGC1a, which in turn activates PPARα to upregulate FAO genes. Dietary supplementation with fish oil containing omega-3 fatty acids, natural PPARα ligands, fully restores quiescence of p53-deficient NSCs and delays tumor initiation in a glioblastoma mouse model. Thus, diet can silence glioblastoma driver mutations, with important implications for cancer prevention.

وصف الملف: text

العلاقة: https://discovery.ucl.ac.uk/id/eprint/10168865/1/1-s2.0-S153458072300148X-main.pdfTest; https://discovery.ucl.ac.uk/id/eprint/10168865Test/

الإتاحة: https://discovery.ucl.ac.uk/id/eprint/10168865/1/1-s2.0-S153458072300148X-main.pdfTest
https://discovery.ucl.ac.uk/id/eprint/10168865Test/

المؤلفون: Ramaprasad, Abhinay, Burda, Paul-Christian, Koussis, Konstantinos, Thomas, James A, Pietsch, Emma, Calvani, Enrica, Howell, Steven A, MacRae, James I, Snijders, Ambrosius P, Gilberger, Tim-Wolf, Blackman, Michael J

المساهمون: Kafsack, Bjorn FC

الوصف: Malaria parasite release (egress) from host red blood cells involves parasite-mediated membrane poration and rupture, thought to involve membrane-lytic effector molecules such as perforin-like proteins and/or phospholipases. With the aim of identifying these effectors, we disrupted the expression of two Plasmodium falciparum perforin-like proteins simultaneously and showed that they have no essential roles during blood stage egress. Proteomic profiling of parasite proteins discharged into the parasitophorous vacuole (PV) just prior to egress detected the presence in the PV of a lecithin:cholesterol acyltransferase (LCAT; PF3D7_0629300). Conditional ablation of LCAT resulted in abnormal egress and a reduced replication rate. Lipidomic profiles of LCAT-null parasites showed drastic changes in several phosphatidylserine and acylphosphatidylglycerol species during egress. We thus show that, in addition to its previously demonstrated role in liver stage merozoite egress, LCAT is required to facilitate efficient egress in asexual blood stage malaria parasites.

وصف الملف: text

العلاقة: https://researchonline.lshtm.ac.uk/id/eprint/4670733/1/Ramaprasad-etal-2023-A-malaria-parasite-phospholipase-facilitates-efficient-asexual-blood-stage-egress.pdfTest; Ramaprasad, Abhinay; Burda, Paul-Christian; Koussis, Konstantinos; Thomas, James A ; Pietsch, Emma; Calvani, Enrica; Howell, Steven A; MacRae, James I; Snijders, Ambrosius P; Gilberger, Tim-Wolf; +1 more. Blackman, Michael J ; (2023) A malaria parasite phospholipase facilitates efficient asexual blood stage egress. PLoS pathogens, 19 (6). e1011449-. ISSN 1553-7366 DOI: https://doi.org/10.1371/journal.ppat.1011449Test

الإتاحة: https://doi.org/10.1371/journal.ppat.1011449Test
https://researchonline.lshtm.ac.uk/id/eprint/4670733Test/
https://researchonline.lshtm.ac.uk/id/eprint/4670733/1/Ramaprasad-etal-2023-A-malaria-parasite-phospholipase-facilitates-efficient-asexual-blood-stage-egress.pdfTest

المؤلفون: de Boer, Leonard L., Vanes, Lesley, Melgrati, Serena, Biggs O’May, Joshua, Hayward, Darryl, Driscoll, Paul C., Day, Jason, Griffiths, Alexander, Magueta, Renata, Morrell, Alexander, MacRae, James I., Köchl, Robert, Tybulewicz, Victor L.J.

المصدر: de Boer , L L , Vanes , L , Melgrati , S , Biggs O’May , J , Hayward , D , Driscoll , P C , Day , J , Griffiths , A , Magueta , R , Morrell , A , MacRae , J I , Köchl , R & Tybulewicz , V L J 2023 , ' T cell migration requires ion and water influx to regulate actin polymerization ' , Nature Communications , vol. 14 , no. 1 , 7844 . https://doi.org/10.1038/s41467-023-43423-8Test

الوصف: Migration of T cells is essential for their ability to mount immune responses. Chemokine-induced T cell migration requires WNK1, a kinase that regulates ion influx into the cell. However, it is not known why ion entry is necessary for T cell movement. Here we show that signaling from the chemokine receptor CCR7 leads to activation of WNK1 and its downstream pathway at the leading edge of migrating CD4 + T cells, resulting in ion influx and water entry by osmosis. We propose that WNK1-induced water entry is required to swell the membrane at the leading edge, generating space into which actin filaments can polymerize, thereby facilitating forward movement of the cell. Given the broad expression of WNK1 pathway proteins, our study suggests that ion and water influx are likely to be essential for migration in many cell types, including leukocytes and metastatic tumor cells.

الإتاحة: https://doi.org/10.1038/s41467-023-43423-8Test
https://kclpure.kcl.ac.uk/portal/en/publications/bd1510c8-8e9a-4e6e-9293-35ece3eca6bbTest
http://www.scopus.com/inward/record.url?scp=85178940164&partnerID=8YFLogxKTest

المؤلفون: Rzechorzek, Neil J., Kunzelmann, Simone, Purkiss, Andrew G., Silva Dos Santos, Mariana, MacRae, James I., Taylor, Ian A., Fugger, Kasper, West, Stephen C.

المصدر: Nature Communications ; volume 14, issue 1 ; ISSN 2041-1723

مصطلحات موضوعية: General Physics and Astronomy, General Biochemistry, Genetics and Molecular Biology, General Chemistry, Multidisciplinary

الوصف: Poly(ADP-ribose) polymerase (PARP) inhibitors are used in the clinic to treat BRCA -deficient breast, ovarian and prostate cancers. As their efficacy is potentiated by loss of the nucleotide salvage factor DNPH1 there is considerable interest in the development of highly specific small molecule DNPH1 inhibitors. Here, we present X-ray crystal structures of dimeric DNPH1 bound to its substrate hydroxymethyl deoxyuridine monophosphate (hmdUMP). Direct interaction with the hydroxymethyl group is important for substrate positioning, while conserved residues surrounding the base facilitate target discrimination. Glycosidic bond cleavage is driven by a conserved catalytic triad and proceeds via a two-step mechanism involving formation and subsequent disruption of a covalent glycosyl-enzyme intermediate. Mutation of a previously uncharacterised yet conserved glutamate traps the intermediate in the active site, demonstrating its role in the hydrolytic step. These observations define the enzyme’s catalytic site and mechanism of hydrolysis, and provide important insights for inhibitor discovery.

الإتاحة: https://doi.org/10.1038/s41467-023-42544-4Test
https://www.nature.com/articles/s41467-023-42544-4.pdfTest
https://www.nature.com/articles/s41467-023-42544-4Test

المؤلفون: Ocasio, Cory A., Baggelaar, Marc, Sipthorp, James, Losada de la Lastra, Ana, Volarić, Jana, Soudy, Christelle, Storck, Elisabeth M., Palma-Duran, Susana A., MacRae, James I., Tomic, Goran, Carr, Lotte, Downward, Julian, S. Eggert, Ulrike, Tate, Edward W.

مصطلحات موضوعية: ZDHHC, Palmitoylation, Proteomics, Chemical genetics

الوصف: The 23 human ZDHHC S-acyltransferases catalyze long-chain S-acylation at cysteine residues across an extensive network of hundreds of proteins important for normal physiology or dysregulated in disease. Here we present a technology platform to directly map the protein substrates of a specific ZDHHC for the first time at the whole proteome level, in intact cells. Structure-guided engineering of paired ZDHHC ‘hole’ mutants and ‘bumped’ chemically tagged fatty acid probes enabled probe transfer to specific protein substrates with excellent selectivity over wild type ZDHHCs. Chemical genetic systems were exemplified for five ZDHHCs (3, 7, 11, 15 and 20), and applied to generate the first de novo ZDHHC substrate profiles, identifying >300 unique and shared substrates across multiple cell lines and Sacylation sites for novel functionally diverse substrates. We expect that this powerful and versatile platform will open a new window on S-acylation biology for a wide range of models and organisms.

وصف الملف: application/pdf

العلاقة: https://dspace.library.uu.nl/handle/1874/436065Test

الإتاحة: https://dspace.library.uu.nl/handle/1874/436065Test

المؤلفون: Grimm, Fiona, Asuaje, Agustín, Jain, Aakriti, Silva dos Santos, Mariana, Kleinjung, Jens, Nunes, Patrícia M, Gehrig, Stefanie, Fets, Louise, Darici, Salihanur, MacRae, James I, Anastasiou, Dimitrios

المصدر: EMBO Journal; Apr2024, Vol. 43 Issue 8, p1545-1569, 25p

مصطلحات موضوعية: CELL survival, CANCER cells, GLYCOLYSIS, HYPOXEMIA, MALATE dehydrogenase, NAD (Coenzyme), HYPOXIA-inducible factor 1, BREAST

مستخلص: Adaptation to chronic hypoxia occurs through changes in protein expression, which are controlled by hypoxia-inducible factor 1α (HIF1α) and are necessary for cancer cell survival. However, the mechanisms that enable cancer cells to adapt in early hypoxia, before the HIF1α-mediated transcription programme is fully established, remain poorly understood. Here we show in human breast cancer cells, that within 3 h of hypoxia exposure, glycolytic flux increases in a HIF1α-independent manner but is limited by NAD⁺ availability. Glycolytic ATP maintenance and cell survival in early hypoxia rely on reserve lactate dehydrogenase A capacity as well as the activity of glutamate-oxoglutarate transaminase 1 (GOT1), an enzyme that fuels malate dehydrogenase 1 (MDH1)-derived NAD⁺. In addition, GOT1 maintains low α-ketoglutarate levels, thereby limiting prolyl hydroxylase activity to promote HIF1α stabilisation in early hypoxia and enable robust HIF1α target gene expression in later hypoxia. Our findings reveal that, in normoxia, multiple enzyme systems maintain cells in a primed state ready to support increased glycolysis and HIF1α stabilisation upon oxygen limitation, until other adaptive processes that require more time are fully established. Synopsis: Long term survival of cancer cells in hypoxia is dependent upon HIF1α activity for energy production. However, it is unclear how cells survive before HIF1α-transcribed genes are robustly expressed. This study investigated the early mechanism of human cancer cell survival upon hypoxia exposure. Shortly upon exposure to hypoxia, human breast cancer cells have increased glycolysis independent of HIF1α and without detectable changes in glycolytic enzyme expression. This glycolytic boost relies on NAD⁺ that is provided by reserve lactate dehydrogenase A (LDHA) capacity and malate dehydrogenase 1 (MDH1) activity fuelled by glutamate-oxoglutarate transaminase 1 (GOT1). However, NAD⁺ is still limiting for maximal flux through ATP-producing lower glycolysis, as indicated by efflux of glucose carbons to α-glycerophosphate (α-GP). Although MDH1 flux does not increase, combined inhibition of GOT1 and LDHA results in attenuated ATP production and cell survival. Consumption of α-ketoglutarate by GOT1 also limits prolyl hydroxylase activity and enables HIF1α stabilisation for long term hypoxic responses. In early hypoxia, NAD⁺ required for glycolytic ATP production is provided by lactate dehydrogenase A and GOT1-fuelled malate dehydrogenase 1 activities before HIF1α-mediated cell survival mechanisms are established. [ABSTRACT FROM AUTHOR]

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المؤلفون: Bussi, Claudio, Heunis, Tiaan, Pellegrino, Enrica, Bernard, Elliott M., Bah, Nourdine, Santos, Mariana Silva dos, Santucci, Pierre, Aylan, Beren, Rodgers, Angela, Fearns, Antony, Mitschke, Julia, Moore, Christopher, MacRae, James I., Greco, Maria, Reinheckel, Thomas, Trost, Matthias, Gutierrez, Maximiliano G.

المصدر: Nature communications. - 13 (2022) , 7338, ISSN: 2041-1723

الوصف: Transient lysosomal damage after infection with cytosolic pathogens or silica crystals uptake results in protease leakage. Whether limited leakage of lysosomal contents into the cytosol affects the function of cytoplasmic organelles is unknown. Here, we show that sterile and non-sterile lysosomal damage triggers a cell death independent proteolytic remodelling of the mitochondrial proteome in macrophages. Mitochondrial metabolic reprogramming required leakage of lysosomal cathepsins and was independent of mitophagy, mitoproteases and proteasome degradation. In an in vivo mouse model of endomembrane damage, live lung macrophages that internalised crystals displayed impaired mitochondrial function. Single-cell RNA-sequencing revealed that lysosomal damage skewed metabolic and immune responses in alveolar macrophages subsets with increased lysosomal content. Functionally, drug modulation of macrophage metabolism impacted host responses to Mycobacterium tuberculosis infection in an endomembrane damage dependent way. This work uncovers an inter-organelle communication pathway, providing a general mechanism by which macrophages undergo mitochondrial metabolic reprograming after endomembrane damage.

وصف الملف: pdf

العلاقة: https://freidok.uni-freiburg.de/data/231625Test

الإتاحة: https://doi.org/10.1038/s41467-022-34632-8Test
https://freidok.uni-freiburg.de/data/231625Test
https://nbn-resolving.org/urn:nbn:de:bsz:25-freidok-2316256Test
https://freidok.uni-freiburg.de/dnb/download/231625Test