The P-body component USP52/PAN2 is a novel regulator of HIF1A mRNA stability

HIF1A (hypoxia-inducible factor 1α) is the master regulator of the cellular response to hypoxia and is implicated in cancer progression. Whereas the regulation of HIF1A protein in response to oxygen is well characterized, less is known about the fate of HIF1A mRNA. In the present study, we have identified the pseudo-DUB (deubiquitinating enzyme)/deadenylase USP52 (ubiquitin-specific protease 52)/PAN2 [poly(A) nuclease 2] as an important regulator of the HIF1A-mediated hypoxic response. Depletion of USP52 reduced HIF1A mRNA and protein levels and resulted in reduced expression of HIF1A-regulated hypoxic targets due to a 3′-UTR (untranslated region)-dependent poly(A)-tail-length-independent destabilization in HIF1A mRNA. MS analysis revealed an association of USP52 with several P-body (processing body) components and we confirmed further that USP52 protein and HIF1A mRNA co-localized with cytoplasmic P-bodies. Importantly, P-body dispersal by knockdown of GW182 or LSM1 resulted in a reduction of HIF1A mRNA levels. These data uncover a novel role for P-bodies in regulating HIF1A mRNA stability, and demonstrate that USP52 is a key component of P-bodies required to prevent HIF1A mRNA degradation.

[1]  P. Lehner,et al.  The RNA-binding E3 ubiquitin ligase MEX-3C links ubiquitination with MHC-I mRNA degradation , 2012, The EMBO journal.

[2]  Steffen Schmidt,et al.  The Caenorhabditis elegans GW182 protein AIN-1 interacts with PAB-1 and subunits of the PAN2-PAN3 and CCR4-NOT deadenylase complexes , 2012, Nucleic acids research.

[3]  Brian Keith,et al.  HIF1α and HIF2α: sibling rivalry in hypoxic tumour growth and progression , 2011, Nature Reviews Cancer.

[4]  W. Filipowicz,et al.  miRNA repression involves GW182-mediated recruitment of CCR4–NOT through conserved W-containing motifs , 2011, Nature Structural &Molecular Biology.

[5]  S. Rocha,et al.  The chromatin remodeler ISWI regulates the cellular response to hypoxia: role of FIH , 2011, Molecular biology of the cell.

[6]  N. Sonenberg,et al.  miRNA-mediated deadenylation is orchestrated by GW182 through two conserved motifs that interact with CCR4–NOT , 2011, Nature Structural &Molecular Biology.

[7]  Joerg E Braun,et al.  GW182 proteins directly recruit cytoplasmic deadenylase complexes to miRNA targets. , 2011, Molecular cell.

[8]  Françoise Bono,et al.  Hypoxia-inducible factor-1α mRNA: a new target for destabilization by tristetraprolin in endothelial cells , 2011, Molecular biology of the cell.

[9]  E. Kondo,et al.  MicroRNA 130 family regulates the hypoxia response signal through the P-body protein DDX6 , 2011, Nucleic acids research.

[10]  A. Shyu,et al.  Mechanisms of deadenylation‐dependent decay , 2011, Wiley interdisciplinary reviews. RNA.

[11]  P. Giannakakou,et al.  Microtubule disruption targets HIF-1α mRNA to cytoplasmic P-bodies for translational repression , 2011, The Journal of cell biology.

[12]  A. Bertolin,et al.  Drosophila Genome-Wide RNAi Screen Identifies Multiple Regulators of HIF–Dependent Transcription in Hypoxia , 2010, PLoS genetics.

[13]  G. Semenza,et al.  Hsp70 and CHIP Selectively Mediate Ubiquitination and Degradation of Hypoxia-inducible Factor (HIF)-1α but Not HIF-2α* , 2009, The Journal of Biological Chemistry.

[14]  S. Gygi,et al.  Defining the Human Deubiquitinating Enzyme Interaction Landscape , 2009, Cell.

[15]  Amanda Doucette,et al.  An inhibitor of NEDD8-activating enzyme as a new approach to treat cancer , 2009, Nature.

[16]  G. Powis,et al.  Hypoxia-Associated Factor, a Novel E3-Ubiquitin Ligase, Binds and Ubiquitinates Hypoxia-Inducible Factor 1α, Leading to Its Oxygen-Independent Degradation , 2008, Molecular and Cellular Biology.

[17]  S. Rocha,et al.  Regulation of gene expression by hypoxia. , 2008, The Biochemical journal.

[18]  A. Shyu,et al.  Deadenylation is prerequisite for P-body formation and mRNA decay in mammalian cells , 2008, The Journal of cell biology.

[19]  J. Lykke-Andersen,et al.  TTP and BRF proteins nucleate processing body formation to silence mRNAs with AU-rich elements. , 2007, Genes & development.

[20]  R. Cole,et al.  RACK1 Competes with HSP90 for Binding to HIF-1α and is Required for O2-independent and HSP90 Inhibitor-induced Degradation of HIF-1α , 2007 .

[21]  George A. Calin,et al.  A MicroRNA Signature of Hypoxia , 2006, Molecular and Cellular Biology.

[22]  Tsung-Cheng Chang,et al.  Concerted action of poly(A) nucleases and decapping enzyme in mammalian mRNA turnover , 2005, Nature Structural &Molecular Biology.

[23]  Randal J. Kaufman,et al.  Stress granules and processing bodies are dynamically linked sites of mRNP remodeling , 2005, The Journal of cell biology.

[24]  E. Chan,et al.  GW182 is critical for the stability of GW bodies expressed during the cell cycle and cell proliferation , 2004, Journal of Cell Science.

[25]  A. Zou,et al.  Androgens regulate the binding of endogenous HuR to the AU-rich 3'UTRs of HIF-1alpha and EGF mRNA. , 2004, Biochemical and biophysical research communications.

[26]  A. Jacobson,et al.  Positive and Negative Regulation of Poly(A) Nuclease , 2004, Molecular and Cellular Biology.

[27]  M. Matthay,et al.  Prolonged Hypoxia Differentially Regulates Hypoxia-inducible Factor (HIF)-1α and HIF-2α Expression in Lung Epithelial Cells , 2004, Journal of Biological Chemistry.

[28]  Víctor Quesada,et al.  Cloning and enzymatic analysis of 22 novel human ubiquitin-specific proteases. , 2004, Biochemical and biophysical research communications.

[29]  S. Hoshino,et al.  Identification of a Human Cytoplasmic Poly(A) Nuclease Complex Stimulated by Poly(A)-binding Protein* , 2004, Journal of Biological Chemistry.

[30]  Brian Keith,et al.  Differential Roles of Hypoxia-Inducible Factor 1α (HIF-1α) and HIF-2α in Hypoxic Gene Regulation , 2003, Molecular and Cellular Biology.

[31]  C. Vaché,et al.  Natural antisense transcripts of hypoxia-inducible factor 1alpha are detected in different normal and tumour human tissues. , 2002, Gene.

[32]  Michael I. Wilson,et al.  C. elegans EGL-9 and Mammalian Homologs Define a Family of Dioxygenases that Regulate HIF by Prolyl Hydroxylation , 2001, Cell.

[33]  R. Parker,et al.  The Transcription Factor Associated Ccr4 and Caf1 Proteins Are Components of the Major Cytoplasmic mRNA Deadenylase in Saccharomyces cerevisiae , 2001, Cell.

[34]  C. Wykoff,et al.  The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis , 1999, Nature.

[35]  Jill K Thompson,et al.  Nuclear Retention of IκBα Protects It from Signal-induced Degradation and Inhibits Nuclear Factor κB Transcriptional Activation* , 1999, The Journal of Biological Chemistry.

[36]  J. Caro,et al.  Hypoxia-inducible factor 1alpha (HIF-1alpha) protein is rapidly degraded by the ubiquitin-proteasome system under normoxic conditions. Its stabilization by hypoxia depends on redox-induced changes. , 1997, The Journal of biological chemistry.

[37]  A. Sachs,et al.  PAN3 encodes a subunit of the Pab1p-dependent poly(A) nuclease in Saccharomyces cerevisiae , 1996, Molecular and cellular biology.

[38]  M. Rieger,et al.  The Yeast Pan2 Protein Is Required for Poly(A)-binding Protein-stimulated Poly(A)-nuclease Activity * , 1996, The Journal of Biological Chemistry.

[39]  K. Abromeit.  Music Received , 2023, Notes.

[40]  Young Il Yeom,et al.  Tristetraprolin regulates the stability of HIF-1alpha mRNA during prolonged hypoxia. , 2010, Biochemical and biophysical research communications.

[41]  E. Izaurralde,et al.  P bodies: at the crossroads of post-transcriptional pathways , 2007, Nature Reviews Molecular Cell Biology.