CLUH controls astrin-1 expression to couple mitochondrial metabolism to cell cycle progression

Proliferating cells undergo metabolic changes in synchrony with cell cycle progression and cell division. Mitochondria provide fuel, metabolites, and ATP during different phases of the cell cycle, however it is not completely understood how mitochondrial function and the cell cycle are coordinated. CLUH is a post-transcriptional regulator of mRNAs encoding mitochondrial proteins involved in oxidative phosphorylation and several metabolic pathways. Here, we show a role of CLUH in regulating the expression of astrin, which is involved in metaphase to anaphase progression, centrosome integrity, and mTORC1 inhibition. We find that CLUH binds both the SPAG5 mRNA and its product astrin, and controls the synthesis and the stability of the full-length astrin-1 isoform. We show that CLUH interacts with astrin-1 specifically during interphase. Astrin-depleted cells show mTORC1 hyperactivation and enhanced anabolism. On the other hand, cells lacking CLUH show decreased astrin levels and increased mTORC1 signaling, but cannot sustain anaplerotic and anabolic pathways. In absence of CLUH, cells fail to grow during G1, and progress faster through the cell cycle, indicating dysregulated matching of growth, metabolism and cell cycling. Our data reveal a role of CLUH in coupling growth signaling pathways and mitochondrial metabolism with cell cycle progression.

[1]  Johana Chicher,et al.  The interactome of CLUH reveals its association to SPAG5 and its co-translational proximity to mitochondrial proteins , 2022, BMC Biology.

[2]  U. Gruneberg,et al.  The association of Plk1 with the astrin–kinastrin complex promotes formation and maintenance of a metaphase plate , 2020, Journal of Cell Science.

[3]  Ou Sha,et al.  Astrin: A Key Player in Mitosis and Cancer , 2020, Frontiers in Cell and Developmental Biology.

[4]  E. Rugarli,et al.  CLUH granules coordinate translation of mitochondrial proteins with mTORC1 signaling and mitophagy , 2020, The EMBO journal.

[5]  L. Fajas,et al.  Cell cycle regulators in cancer cell metabolism. , 2020, Biochimica et biophysica acta. Molecular basis of disease.

[6]  Nicola Zamboni,et al.  Lipid signalling drives proteolytic rewiring of mitochondria by YME1L , 2019, Nature.

[7]  B. Habermann,et al.  mitoXplorer, a visual data mining platform to systematically analyze and visualize mitochondrial expression dynamics and mutations , 2019, bioRxiv.

[8]  Wentao Yang,et al.  SPAG5 upregulation contributes to enhanced c-MYC transcriptional activity via interaction with c-MYC binding protein in triple-negative breast cancer , 2019, Journal of Hematology & Oncology.

[9]  E. Rugarli,et al.  A concert of RNA-binding proteins coordinates mitochondrial function , 2018, Critical reviews in biochemistry and molecular biology.

[10]  Ji Jing,et al.  Sperm-Associated Antigen 5 Expression Is Increased in Hepatocellular Carcinoma and Indicates Poor Prognosis , 2018, Medical science monitor : international medical journal of experimental and clinical research.

[11]  Marcus Krüger,et al.  Instant Clue: A Software Suite for Interactive Data Visualization and Analysis , 2018, Scientific Reports.

[12]  Pär Nordlund,et al.  Modulation of Protein-Interaction States through the Cell Cycle , 2018, Cell.

[13]  R. Deberardinis,et al.  Loss of a Negative Regulator of mTORC1 Induces Aerobic Glycolysis and Altered Fiber Composition in Skeletal Muscle , 2018, Cell reports.

[14]  S. Mohammed,et al.  Doxorubicin-induced DNA Damage Causes Extensive Ubiquitination of Ribosomal Proteins Associated with a Decrease in Protein Translation* , 2018, Molecular & Cellular Proteomics.

[15]  T. Shlomi,et al.  Temporal fluxomics reveals oscillations in TCA cycle flux throughout the mammalian cell cycle , 2017, Molecular systems biology.

[16]  H. McBride,et al.  mTOR Controls Mitochondrial Dynamics and Cell Survival via MTFP1. , 2017, Molecular cell.

[17]  V. Velagapudi,et al.  mTORC1 Regulates Mitochondrial Integrated Stress Response and Mitochondrial Myopathy Progression. , 2017, Cell metabolism.

[18]  Evelyn Rampler,et al.  Anion-Exchange Chromatography Coupled to High-Resolution Mass Spectrometry: A Powerful Tool for Merging Targeted and Non-targeted Metabolomics. , 2017, Analytical chemistry.

[19]  Kuan-Chung Su,et al.  Astrin-SKAP complex reconstitution reveals its kinetochore interaction with microtubule-bound Ndc80 , 2017, bioRxiv.

[20]  D. Logan,et al.  CLUH couples mitochondrial distribution to the energetic and metabolic status , 2017, Journal of Cell Science.

[21]  E. Rugarli,et al.  CLUH regulates mitochondrial metabolism by controlling translation and decay of target mRNAs , 2017, The Journal of cell biology.

[22]  Iain M Cheeseman,et al.  Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the Diversity of p53-Dependent Responses to Cell-Cycle Defects. , 2017, Developmental cell.

[23]  M. Malumbres,et al.  Fueling the Cell Division Cycle. , 2017, Trends in cell biology.

[24]  Jüergen Cox,et al.  The MaxQuant computational platform for mass spectrometry-based shotgun proteomics , 2016, Nature Protocols.

[25]  G. Ball,et al.  SPAG5 as a prognostic biomarker and chemotherapy sensitivity predictor in breast cancer: a retrospective, integrated genomic, transcriptomic, and protein analysis. , 2016, The Lancet. Oncology.

[26]  F. Bertucci,et al.  SPAG5: the ultimate marker of proliferation in early breast cancer? , 2016, The Lancet. Oncology.

[27]  R. Wek,et al.  Upstream Open Reading Frames Differentially Regulate Gene-specific Translation in the Integrated Stress Response* , 2016, The Journal of Biological Chemistry.

[28]  Marco Y. Hein,et al.  The Perseus computational platform for comprehensive analysis of (prote)omics data , 2016, Nature Methods.

[29]  Chang-Young Jang,et al.  Phosphorylation of Astrin Regulates Its Kinetochore Function* , 2016, The Journal of Biological Chemistry.

[30]  O. Mabrouk,et al.  Benzoyl chloride derivatization with liquid chromatography-mass spectrometry for targeted metabolomics of neurochemicals in biological samples. , 2016, Journal of chromatography. A.

[31]  D. Page,et al.  A mitotic SKAP isoform regulates spindle positioning at astral microtubule plus ends , 2016, The Journal of cell biology.

[32]  Andrew D. Rouillard,et al.  Enrichr: a comprehensive gene set enrichment analysis web server 2016 update , 2016, Nucleic Acids Res..

[33]  F. Herzog,et al.  Molecular requirements for the inter-subunit interaction and kinetochore recruitment of SKAP and Astrin , 2016, Nature Communications.

[34]  J. Whitelegge,et al.  The X-Linked-Intellectual-Disability-Associated Ubiquitin Ligase Mid2 Interacts with Astrin and Regulates Astrin Levels to Promote Cell Division. , 2016, Cell reports.

[35]  C. Walsh,et al.  Centriolar satellites assemble centrosomal microcephaly proteins to recruit CDK2 and promote centriole duplication , 2015, eLife.

[36]  J. Yun,et al.  SPAG5 upregulation predicts poor prognosis in cervical cancer patients and alters sensitivity to taxol treatment via the mTOR signaling pathway , 2015, Cell Death and Disease.

[37]  Oliver M. Bernhardt,et al.  Extending the Limits of Quantitative Proteome Profiling with Data-Independent Acquisition and Application to Acetaminophen-Treated Three-Dimensional Liver Microtissues* , 2015, Molecular & Cellular Proteomics.

[38]  Josip S. Herman,et al.  Cell cycle–dependent regulation of mitochondrial preprotein translocase , 2014, Science.

[39]  E. Rugarli,et al.  CLUH regulates mitochondrial biogenesis by binding mRNAs of nuclear-encoded mitochondrial proteins , 2014, The Journal of cell biology.

[40]  J. Chen,et al.  The mitosis-regulating and protein-protein interaction activities of astrin are controlled by aurora-A-induced phosphorylation. , 2014, American journal of physiology. Cell physiology.

[41]  F. He,et al.  Cyclin B1/Cdk1 coordinates mitochondrial respiration for cell-cycle G2/M progression. , 2014, Developmental cell.

[42]  E. Cuyás,et al.  Cell cycle regulation by the nutrient-sensing mammalian target of rapamycin (mTOR) pathway. , 2014, Methods in molecular biology.

[43]  In Hye Lee,et al.  Metabolic regulation of the cell cycle. , 2013, Current opinion in cell biology.

[44]  N. Sonenberg,et al.  mTORC1 controls mitochondrial activity and biogenesis through 4E-BP-dependent translational regulation. , 2013, Cell metabolism.

[45]  M. Hall,et al.  Inhibition of mTORC1 by Astrin and Stress Granules Prevents Apoptosis in Cancer Cells , 2013, Cell.

[46]  Shabaz Mohammed,et al.  Comparative Phosphoproteomic Analysis of Checkpoint Recovery Identifies New Regulators of the DNA Damage Response , 2013, Science Signaling.

[47]  E. Chen,et al.  Enrichr: interactive and collaborative HTML5 gene list enrichment analysis tool , 2013, BMC Bioinformatics.

[48]  F. Barr,et al.  The astrin–kinastrin/SKAP complex localizes to microtubule plus ends and facilitates chromosome alignment , 2011, The Journal of cell biology.

[49]  D. Compton,et al.  CLASP1, astrin and Kif2b form a molecular switch that regulates kinetochore‐microtubule dynamics to promote mitotic progression and fidelity , 2010, The EMBO journal.

[50]  T. Hori,et al.  Aurora B kinase controls the targeting of the Astrin–SKAP complex to bioriented kinetochores , 2010, The Journal of cell biology.

[51]  S. Kornbluth,et al.  The engine driving the ship: metabolic steering of cell proliferation and death , 2010, Nature reviews. Molecular cell biology.

[52]  S. Brunak,et al.  Quantitative Phosphoproteomics Reveals Widespread Full Phosphorylation Site Occupancy During Mitosis , 2010, Science Signaling.

[53]  Badrinath Roysam,et al.  A hyperfused mitochondrial state achieved at G1–S regulates cyclin E buildup and entry into S phase , 2009, Proceedings of the National Academy of Sciences.

[54]  Chi-Ying F. Huang,et al.  Glycogen Synthase Kinase 3β Interacts with and Phosphorylates the Spindle-associated Protein Astrin* , 2008, Journal of Biological Chemistry.

[55]  R. Deberardinis,et al.  The biology of cancer: metabolic reprogramming fuels cell growth and proliferation. , 2008, Cell metabolism.

[56]  E. Nigg,et al.  Astrin is required for the maintenance of sister chromatid cohesion and centrosome integrity , 2007, The Journal of cell biology.

[57]  Chi-Ying F. Huang,et al.  hNinein is required for targeting spindle-associated protein Astrin to the centrosome during the S and G2 phases. , 2007, Experimental cell research.

[58]  M. Mann,et al.  Protocol for micro-purification, enrichment, pre-fractionation and storage of peptides for proteomics using StageTips , 2007, Nature Protocols.

[59]  E. Rugarli,et al.  Variable and Tissue-Specific Subunit Composition of Mitochondrial m-AAA Protease Complexes Linked to Hereditary Spastic Paraplegia , 2006, Molecular and Cellular Biology.

[60]  Utpal Banerjee,et al.  Mitochondrial regulation of cell cycle progression during development as revealed by the tenured mutation in Drosophila. , 2005, Developmental cell.

[61]  Jens Gruber,et al.  The mitotic-spindle-associated protein astrin is essential for progression through mitosis , 2002, Journal of Cell Science.

[62]  Yuh Cheng Yang,et al.  Cloning and characterization of hMAP126, a new member of mitotic spindle-associated proteins. , 2001, Biochemical and biophysical research communications.

[63]  D. Morris,et al.  Upstream Open Reading Frames as Regulators of mRNA Translation , 2000, Molecular and Cellular Biology.

[64]  Benzoyl chloride. , 1982, IARC monographs on the evaluation of the carcinogenic risk of chemicals to humans.