GTPase Activating Protein (Sh3 Domain) Binding Protein 1 Regulates the Processing of MicroRNA-1 during Cardiac Hypertrophy

Background MicroRNAs (miR) are small, posttranscriptional regulators, expressed as part of a longer primary transcript, following which they undergo nuclear and cytoplasmic processing by Drosha and Dicer, respectively, to form the functional mature ~20mer that gets incorporated into the silencing complex. Others and we have shown that mature miR-1 levels decrease with pressure-induced cardiac hypertrophy, however, there is little or no change in the primary transcript encompassing miR-1 stem-loop, suggesting critical regulatory step in microRNA processing. The objective of this study was to investigate the underlying mechanisms regulating miR-1 expression in cardiomyocytes. Results Here we report that GTPase–activating protein (SH3 domain) binding protein 1 (G3bp1), an endoribonuclease regulates miR-1 processing in cardiomyocytes. G3bp1 is upregulated during cardiac hypertrophy and restricts miR-1 processing by binding to its consensus sequence in the pre-miR-1-2 stem-loop. In accordance, exogenous G3bp1 is sufficient to reduce miR-1 levels, along with derepression of miR-1 targets; General transcription factor IIB (Gtf2b), cyclin dependent factor 9 (Cdk9) and eukaryotic initiation factor 4E (Eif4e). While Cdk9 and Gtf2b are essential for transcription, Eif4e is required for translation. Thus, downregulation of miR-1 is necessary for increase in these molecules. Similar to miR-1 knockdown, G3bp1 overexpression is not sufficient for development of cardiac hypertrophy. Conversely, knockdown of G3bp1 in hypertrophying cardiomyocytes inhibited downregulation of miR-1 and upregulation of its targets along with restricted hypertrophy, suggesting that G3bp1 is necessary for development of cardiac hypertrophy. These results indicate that G3bp1-mediated inhibition of miR-1 processing with growth stimulation results in decrease in mature miR-1 and, thereby, an increase of its targets, which play fundamental roles in the development of hypertrophy. Conclusion G3bp1 posttranscriptionally regulates miRNA-1 processing in the heart, and G3bp1 mediated downregulation of mature miRNA-1 levels is required for the derepression of its targets and increase in gene expression during cardiac hypertrophy.

[1]  J. Tazi,et al.  Control of Fetal Growth and Neonatal Survival by the RasGAP-Associated Endoribonuclease G3BP , 2005, Molecular and Cellular Biology.

[2]  A. Ramos,et al.  Functional and molecular insights into KSRP function in mRNA decay. , 2013, Biochimica et biophysica acta.

[3]  Kotb Abdelmohsen,et al.  Posttranscriptional gene regulation by long noncoding RNA. , 2013, Journal of molecular biology.

[4]  D. Catalucci,et al.  Reciprocal Regulation of MicroRNA-1 and Insulin-Like Growth Factor-1 Signal Transduction Cascade in Cardiac and Skeletal Muscle in Physiological and Pathological Conditions , 2009, Circulation.

[5]  Jian-Fu Chen,et al.  The role of microRNA-1 and microRNA-133 in skeletal muscle proliferation and differentiation , 2006, Nature Genetics.

[6]  C. Burge,et al.  Most mammalian mRNAs are conserved targets of microRNAs. , 2008, Genome research.

[7]  G. Scheper,et al.  Eukaryotic initiation factors-4E and -4F stimulate 5' cap-dependent as well as internal initiation of protein synthesis. , 1992, The Journal of biological chemistry.

[8]  P. Marsden,et al.  Competition and collaboration between RNA‐binding proteins and microRNAs , 2014, Wiley interdisciplinary reviews. RNA.

[9]  Danish Sayed,et al.  Acute Targeting of General Transcription Factor IIB Restricts Cardiac Hypertrophy via Selective Inhibition of Gene Transcription , 2015, Circulation. Heart failure.

[10]  Christer Larsson,et al.  Regulation of PMP22 mRNA by G3BP1 affects cell proliferation in breast cancer cells , 2013, Molecular Cancer.

[11]  Kevin J Luebke,et al.  Faculty Opinions recommendation of The RNA-binding protein KSRP promotes the biogenesis of a subset of microRNAs. , 2009 .

[12]  T. Sun,et al.  Attenuation of microRNA-1 derepresses the cytoskeleton regulatory protein twinfilin-1 to provoke cardiac hypertrophy , 2010, Journal of Cell Science.

[13]  S. Chien,et al.  Regulation of cardiac gene expression during myocardial growth and hypertrophy: molecular studies of an adaptive physiologic response , 1991, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[14]  B. Carabello,et al.  Translational Initiation Factor eIF-4E , 1996, The Journal of Biological Chemistry.

[15]  Sek Won Kong,et al.  Altered microRNA expression in human heart disease. , 2007, Physiological genomics.

[16]  Danish Sayed,et al.  MicroRNA-21 targets Sprouty2 and promotes cellular outgrowths. , 2008, Molecular biology of the cell.

[17]  V. Ambros,et al.  An Extensive Class of Small RNAs in Caenorhabditis elegans , 2001, Science.

[18]  Sanghyuk Lee,et al.  MicroRNA genes are transcribed by RNA polymerase II , 2004, The EMBO journal.

[19]  L. Lim,et al.  MicroRNA targeting specificity in mammals: determinants beyond seed pairing. , 2007, Molecular cell.

[20]  Jian-Fu Chen,et al.  Expression of microRNAs is dynamically regulated during cardiomyocyte hypertrophy. , 2007, Journal of molecular and cellular cardiology.

[21]  C. Poizat,et al.  Epigenetics and Chromatin Remodeling in Adult Cardiomyopathy , 2013, The Journal of pathology.

[22]  J. Mattick,et al.  Characterization of G3BPs: Tissue specific expression, chromosomal localisation and rasGAP120 binding studies , 2001, Journal of cellular biochemistry.

[23]  A. Cutilletta.  Regression of myocardial hypertrophy. II. RNA synthesis and RNA polymerase activity. , 1980, Journal of molecular and cellular cardiology.

[24]  M. Rosenfeld,et al.  The RNA-binding Protein KSRP Promotes the Biogenesis of a Subset of miRNAs , 2016 .

[25]  V. Kim.  MicroRNA biogenesis: coordinated cropping and dicing , 2005, Nature Reviews Molecular Cell Biology.

[26]  R. Zak,et al.  Muscle and non-muscle cell RNA polymerase activity during the development of myocardial hypertrophy. , 1978, Journal of molecular and cellular cardiology.

[27]  J. Mattick,et al.  Identification of a mouse orthologue of the human ras-GAP-SH3-domain binding protein and structural confirmation that these proteins contain an RNA recognition motif. , 1996, Biomedical peptides, proteins & nucleic acids : structure, synthesis & biological activity.

[28]  Danish Sayed,et al.  Transcriptional Regulation Patterns Revealed by High Resolution Chromatin Immunoprecipitation during Cardiac Hypertrophy* , 2012, The Journal of Biological Chemistry.

[29]  T. Cooper,et al.  RNA-binding proteins in heart development. , 2014, Advances in experimental medicine and biology.

[30]  L. Debussche,et al.  A Ras-GTPase-activating protein SH3-domain-binding protein , 1996, Molecular and cellular biology.

[31]  D. Dembélé,et al.  Misregulation of miR-1 processing is associated with heart defects in myotonic dystrophy , 2011, Nature Structural &Molecular Biology.

[32]  S. Guil,et al.  The multifunctional RNA-binding protein hnRNP A1 is required for processing of miR-18a , 2007, Nature Structural &Molecular Biology.

[33]  T. Hla,et al.  Post-transcriptional gene regulation by HuR and microRNAs in angiogenesis , 2014, Current opinion in hematology.

[34]  R. Jaenisch,et al.  Loss of Cardiac microRNA-Mediated Regulation Leads to Dilated Cardiomyopathy and Heart Failure , 2009, Circulation research.

[35]  A. Gingras,et al.  The mRNA 5' cap-binding protein eIF4E and control of cell growth. , 1998, Current opinion in cell biology.

[36]  Danish Sayed,et al.  MicroRNAs Play an Essential Role in the Development of Cardiac Hypertrophy , 2007, Circulation research.

[37]  M. Lerman,et al.  The mechanism of the heart's adaptation to prolonged load and dynamics of RNA synthesis in the myocardium , 1974, Basic Research in Cardiology.

[38]  S. Pikkarainen,et al.  GATA transcription factors in the developing and adult heart. , 2004, Cardiovascular research.

[39]  P. McDermott,et al.  Increased expression of eukaryotic initiation factor 4E during growth of neonatal rat cardiocytes in vitro. , 1998, American journal of physiology. Heart and circulatory physiology.

[40]  E. Olson,et al.  Control of cardiac growth by histone acetylation/deacetylation. , 2005, Circulation research.

[41]  J. Tazi,et al.  RasGAP-Associated Endoribonuclease G3BP: Selective RNA Degradation and Phosphorylation-Dependent Localization , 2001, Molecular and Cellular Biology.

[42]  K. Gunsalus,et al.  Combinatorial microRNA target predictions , 2005, Nature Genetics.

[43]  J. Tazi,et al.  Deficiency of G3BP1, the stress granules assembly factor, results in abnormal synaptic plasticity and calcium homeostasis in neurons , 2013, Journal of neurochemistry.

[44]  A. Ortega,et al.  Human G3BP1 interacts with β-F1-ATPase mRNA and inhibits its translation , 2010, Journal of Cell Science.

[45]  T. Golub,et al.  MicroRNA-1 Negatively Regulates Expression of the Hypertrophy-Associated Calmodulin and Mef2a Genes , 2009, Molecular and Cellular Biology.

[46]  C. Semple,et al.  Posttranscriptional Regulation of miRNAs Harboring Conserved Terminal Loops , 2008, Molecular cell.

[47]  T. Dudnakova,et al.  Methods Molecular Biology , 2016 .

[48]  Danish Sayed,et al.  MicroRNAs in development and disease. , 2011, Physiological reviews.

[49]  M. Abdellatif,et al.  An Alliance between Ras GTPase-activating Protein, Filamin C, and Ras GTPase-activating Protein SH3 Domain-binding Protein Regulates Myocyte Growth* , 2005, Journal of Biological Chemistry.

[50]  L. Lim,et al.  An Abundant Class of Tiny RNAs with Probable Regulatory Roles in Caenorhabditis elegans , 2001, Science.

[51]  V. Ambros,et al.  The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14 , 1993, Cell.

[52]  K. Chébli,et al.  A Novel Phosphorylation-Dependent RNase Activity of GAP-SH3 Binding Protein: a Potential Link between Signal Transduction and RNA Stability , 1998, Molecular and Cellular Biology.

[53]  K. Chébli,et al.  The RasGAP-associated endoribonuclease G3BP assembles stress granules , 2003, The Journal of cell biology.

[54]  C. Burge,et al.  Conserved Seed Pairing, Often Flanked by Adenosines, Indicates that Thousands of Human Genes are MicroRNA Targets , 2005, Cell.

[55]  Cutilletta Af.  Regression of myocardial hypertrophy II. RNA synthesis and RNA polymerase activity , 1980 .

[56]  D. Srivastava,et al.  The RNA-binding Protein TDP-43 Selectively Disrupts MicroRNA-1/206 Incorporation into the RNA-induced Silencing Complex*♦ , 2014, The Journal of Biological Chemistry.

[57]  Masayasu Oie,et al.  Both G3BP1 and G3BP2 contribute to stress granule formation , 2013, Genes to cells : devoted to molecular & cellular mechanisms.

[58]  Haoming Zhang,et al.  miR-16 family induces cell cycle arrest by regulating multiple cell cycle genes , 2008, Nucleic acids research.

[59]  Martin M Matzuk,et al.  A bioinformatics tool for linking gene expression profiling results with public databases of microRNA target predictions. , 2008, RNA.

[60]  Kevin W Eliceiri,et al.  NIH Image to ImageJ: 25 years of image analysis , 2012, Nature Methods.

[61]  C. Croce,et al.  MicroRNA-133 controls cardiac hypertrophy , 2007, Nature Medicine.