The RNA-binding protein HuR (Elavl1) is essential for the B cell antibody response

Post-transcriptional regulation of mRNA by the RNA-binding protein HuR (encoded by Elavl1) is required in B cells for the germinal center reaction and for the production of class-switched antibodies in response to thymus-independent antigens. Transcriptome-wide examination of RNA isoforms and their abundance and translation in HuR-deficient B cells, together with direct measurements of HuR-RNA interactions, revealed that HuR-dependent splicing of mRNA affected hundreds of transcripts, including that encoding dihydrolipoamide S-succinyltransferase (Dlst), a subunit of the 2-oxoglutarate dehydrogenase (α-KGDH) complex. In the absence of HuR, defective mitochondrial metabolism resulted in large amounts of reactive oxygen species and B cell death. Our study shows how post-transcriptional processes control the balance of energy metabolism required for the proliferation and differentiation of B cells.

[1]  T. Chiles,et al.  Protein kinase Cβ is critical for the metabolic switch to glycolysis following B-cell antigen receptor engagement. , 2012, The Biochemical journal.

[2]  N. Rajewsky,et al.  Regulation of the Germinal Center Response by MicroRNA-155 , 2007, Science.

[3]  A. Ramiro,et al.  Regulation of B‐cell development and function by microRNAs , 2013, Immunological reviews.

[4]  E. Vigorito,et al.  Signal transduction through Vav-2 participates in humoral immune responses and B cell maturation , 2001, Nature Immunology.

[5]  V. Aidinis,et al.  The RNA-Binding Protein Elavl1/HuR Is Essential for Placental Branching Morphogenesis and Embryonic Development , 2009, Molecular and Cellular Biology.

[6]  Scott B. Dewell,et al.  Transcriptome-wide Identification of RNA-Binding Protein and MicroRNA Target Sites by PAR-CLIP , 2010, Cell.

[7]  H. Satoh,et al.  Regulation of Reactive Oxygen Species by Atm Is Essential for Proper Response to DNA Double-Strand Breaks in Lymphocytes1 , 2007, The Journal of Immunology.

[8]  U. Klein,et al.  Germinal center B cell maintenance and differentiation are controlled by distinct NF-κB transcription factor subunits , 2014, The Journal of experimental medicine.

[9]  Caiying Guo,et al.  Essential role of the RNA-binding protein HuR in progenitor cell survival in mice. , 2009, The Journal of clinical investigation.

[10]  A J Carlson,et al.  Frederick R. Rickles, MD, FACP, Appointed Executive Director of the Federation of American Societies for Experimental Biology , 2004, Journal of Investigative Medicine.

[11]  R. Gascoyne,et al.  HVCN1 modulates BCR signal strength via regulation of BCR-dependent generation of reactive oxygen species , 2010, Nature Immunology.

[12]  J. Cadet,et al.  Deamination of the radical cation of the base moiety of 2'-deoxycytidine: a theoretical study. , 2008, Chemphyschem : a European journal of chemical physics and physical chemistry.

[13]  E. Clark,et al.  BCR‐induced superoxide negatively regulates B‐cell proliferation and T‐cell‐independent type 2 Ab responses , 2009, European journal of immunology.

[14]  P. Ivanov,et al.  Post‐transcriptional regulatory networks in immunity , 2013, Immunological reviews.

[15]  V. Bunik,et al.  Phosphonate analogues of alpha-ketoglutarate inhibit the activity of the alpha-ketoglutarate dehydrogenase complex isolated from brain and in cultured cells. , 2005, Biochemistry.

[16]  J. Fujii,et al.  B- to plasma-cell terminal differentiation entails oxidative stress and profound reshaping of the antioxidant responses. , 2010, Antioxidants & redox signaling.

[17]  H. Lardy,et al.  Oxidative phosphorylations; rôle of inorganic phosphate and acceptor systems in control of metabolic rates. , 1952, The Journal of biological chemistry.

[18]  M. Beal,et al.  Mice deficient in dihydrolipoyl succinyl transferase show increased vulnerability to mitochondrial toxins , 2009, Neurobiology of Disease.

[19]  Takashi Tsukamoto,et al.  Glucose-independent glutamine metabolism via TCA cycling for proliferation and survival in B cells. , 2012, Cell metabolism.

[20]  H. Anders,et al.  Loss of Roquin induces early death and immune deregulation but not autoimmunity , 2011, The Journal of experimental medicine.

[21]  D. Guertin,et al.  T cell exit from quiescence and differentiation into Th2 cells depend on Raptor-mTORC1-mediated metabolic reprogramming. , 2013, Immunity.

[22]  Bing Zhang,et al.  WebGestalt: an integrated system for exploring gene sets in various biological contexts , 2005, Nucleic Acids Res..

[23]  J. Ule,et al.  iCLIP reveals the function of hnRNP particles in splicing at individual nucleotide resolution , 2010, Nature Structural &Molecular Biology.

[24]  N. Rajewsky,et al.  Transcriptome-wide analysis of regulatory interactions of the RNA-binding protein HuR. , 2011, Molecular cell.

[25]  E. Vigorito,et al.  Noncoding RNA and its associated proteins as regulatory elements of the immune system , 2014, Nature Immunology.

[26]  L. Tretter,et al.  Generation of Reactive Oxygen Species in the Reaction Catalyzed by α-Ketoglutarate Dehydrogenase , 2004, The Journal of Neuroscience.

[27]  A. DeFranco,et al.  Prolonged Production of Reactive Oxygen Species in Response to B Cell Receptor Stimulation Promotes B Cell Activation and Proliferation , 2012, The Journal of Immunology.

[28]  W. Huber,et al.  Detecting differential usage of exons from RNA-seq data , 2012, Genome research.

[29]  L. Biesecker,et al.  Knockout of Slc25a19 causes mitochondrial thiamine pyrophosphate depletion, embryonic lethality, CNS malformations, and anemia , 2006, Proceedings of the National Academy of Sciences.

[30]  T. Buttke,et al.  Redox regulation of programmed cell death in lymphocytes. , 1995, Free radical research.

[31]  B. van Steensel,et al.  Differential Programming of B Cells in AID Deficient Mice , 2013, PloS one.

[32]  P. Bingham,et al.  A strategically designed small molecule attacks alpha-ketoglutarate dehydrogenase in tumor cells through a redox process , 2014, Cancer & metabolism.

[33]  F. Alt,et al.  Impaired B cell development and proliferation in absence of phosphoinositide 3-kinase p85alpha. , 1999, Science.

[34]  M. Pollak,et al.  Alterations in Cellular Energy Metabolism Associated with the Antiproliferative Effects of the ATM Inhibitor KU-55933 and with Metformin , 2012, PloS one.

[35]  Anton J. Enright,et al.  microRNA-155 regulates the generation of immunoglobulin class-switched plasma cells. , 2007, Immunity.

[36]  Dhiraj Kumar,et al.  The Strength of Receptor Signaling Is Centrally Controlled through a Cooperative Loop between Ca2+ and an Oxidant Signal , 2005, Cell.

[37]  Brad T. Sherman,et al.  Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources , 2008, Nature Protocols.

[38]  Q. Zeng,et al.  The RNase III enzyme Dicer is essential for germinal center B-cell formation. , 2012, Blood.

[39]  L. Tretter,et al.  Inhibition of Krebs Cycle Enzymes by Hydrogen Peroxide: A Key Role of α-Ketoglutarate Dehydrogenase in Limiting NADH Production under Oxidative Stress , 2000, The Journal of Neuroscience.

[40]  M. Beal,et al.  The negative impact of α‐ketoglutarate dehydrogenase complex deficiency on matrix substrate‐level phosphorylation , 2013, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[41]  G E Gibson,et al.  α‐ketoglutarate dehydrogenase in alzheimer brains bearing the APP670/671 mutation , 1998 .

[42]  K. Rajewsky,et al.  MYC is essential for the formation and maintenance of germinal centers , 2012, Nature Immunology.

[43]  Uwe Ohler,et al.  Integrative regulatory mapping indicates that the RNA-binding protein HuR couples pre-mRNA processing and mRNA stability. , 2011, Molecular cell.

[44]  M. Reth,et al.  Testing gene function early in the B cell lineage in mb1-cre mice , 2006, Proceedings of the National Academy of Sciences.

[45]  N. Mukherjee,et al.  Control of Thymic T Cell Maturation, Deletion and Egress by the RNA-Binding Protein HuR1 , 2009, The Journal of Immunology.

[46]  Michael Reth,et al.  Hydrogen peroxide as second messenger in lymphocyte activation , 2002, Nature Immunology.

[47]  M. Dimitriou,et al.  Myeloid cell expression of the RNA-binding protein HuR protects mice from pathologic inflammation and colorectal carcinogenesis. , 2012, The Journal of clinical investigation.

[48]  G. Gibson,et al.  Reduction in the E2k Subunit of the α-Ketoglutarate Dehydrogenase Complex Has Effects Independent of Complex Activity* , 2005, Journal of Biological Chemistry.

[49]  S. Nutt,et al.  The genetic network controlling plasma cell differentiation. , 2011, Seminars in immunology.

[50]  HighWire Press.  The journal of neuroscience : the official journal of the Society for Neuroscience. , 1981 .

[51]  W. Huber,et al.  which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. MAnorm: a robust model for quantitative comparison of ChIP-Seq data sets , 2011 .

[52]  V. Bunik,et al.  The 2-Oxoacid Dehydrogenase Complexes in Mitochondria Can Produce Superoxide/Hydrogen Peroxide at Much Higher Rates Than Complex I* , 2014, The Journal of Biological Chemistry.

[53]  D. Green,et al.  Metabolic Reprogramming Is Required for Antibody Production That Is Suppressed in Anergic but Exaggerated in Chronically BAFF-Exposed B Cells , 2014, The Journal of Immunology.

[54]  Chih-Hao Chang,et al.  Fueling Immunity: Insights into Metabolism and Lymphocyte Function , 2013, Science.

[55]  M. Gorospe,et al.  Conditional Knockout of the RNA-Binding Protein HuR in CD4+ T Cells Reveals a Gene Dosage Effect on Cytokine Production , 2014, Molecular medicine.

[56]  C. E. Schrader,et al.  p53 Represses Class Switch Recombination to IgG2a through Its Antioxidant Function , 2010, The Journal of Immunology.

[57]  Manuel D. Díaz-Muñoz,et al.  Cyclooxygenase-2 Deficiency in Macrophages Leads to Defective p110γ PI3K Signaling and Impairs Cell Adhesion and Migration , 2013, The Journal of Immunology.

[58]  V. Bunik,et al.  Phosphonate Analogues of α-Ketoglutarate Inhibit the Activity of the α-Ketoglutarate Dehydrogenase Complex Isolated from Brain and in Cultured Cells† , 2005 .

[59]  J. Grayson,et al.  Antioxidant Treatment Regulates the Humoral Immune Response during Acute Viral Infection , 2012, Journal of Virology.

[60]  M. Shlomchik,et al.  B Cell Receptor Signal Transduction in the GC Is Short-Circuited by High Phosphatase Activity , 2012, Science.

[61]  R. Schneider,et al.  AUF1 is involved in splenic follicular B cell maintenance , 2010, BMC Immunology.