Contrasting effects of two alternative splicing forms of coactivator-associated arginine methyltransferase 1 on thyroid hormone receptor-mediated transcription in Xenopus laevis.

Thyroid hormone receptors (TRs) can repress or activate target genes depending on the absence or presence of thyroid hormone (T3), respectively. This hormone-dependent gene regulation is mediated by the recruitment of corepressors in the absence of T3 and coactivators in its presence. Many TR-interacting coactivators have been characterized in vitro. Among them is coactivator-associated arginine methyltransferase 1 (CARM1), which methylates histone H3. We are interested in investigating the role of CARM1 in TR-mediated gene expression in vivo during postembryonic development by using T3-dependent frog metamorphosis as a model. We first cloned the Xenopus laevis CARM1 and obtained two alternative splicing forms, CARM1a and CARM1b. Both isoforms are expressed throughout metamorphosis, supporting a role for these isoforms during the process. To investigate whether Xenopus CARM1s participate in gene regulation by TRs, transcriptional analysis was conducted in Xenopus oocyte, where the effects of cofactors can be studied in the context of chromatin in vivo. Surprisingly, overexpression of CARM1b had little effect on TR-mediated transcription, whereas CARM1a enhanced gene activation by liganded TR. Chromatin immunoprecipitation assays showed that both endogenous CARM1a and overexpressed CARM1a and b were recruited to the promoter by liganded TR. However, the binding of liganded TR to the target promoter was reduced when CARM1b was overexpressed, accompanied by a slight reduction in histone methylation at the promoter. These results suggest that CARM1 may play a role in TR-mediated transcriptional regulation during frog development and that its function is regulated by alternative splicing.

[1]  L. Burke,et al.  Co‐repressors 2000 , 2000, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[2]  Yunbo Shi Amphibian Metamorphosis: From Morphology to Molecular Biology , 1999 .

[3]  M. Lazar,et al.  The mechanism of action of thyroid hormones. , 2000, Annual review of physiology.

[4]  N. Marsh-Armstrong,et al.  Diverse developmental programs of Xenopus laevis metamorphosis are inhibited by a dominant negative thyroid hormone receptor , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[5]  D. Aswad,et al.  Methylation of histone H3 by coactivator-associated arginine methyltransferase 1. , 2001, Biochemistry.

[6]  N. McKenna,et al.  Nuclear Receptors, Coregulators, Ligands, and Selective Receptor Modulators , 2001, Annals of the New York Academy of Sciences.

[7]  Yunbo Shi,et al.  A Dominant-Negative Thyroid Hormone Receptor Blocks Amphibian Metamorphosis by Retaining Corepressors at Target Genes , 2003, Molecular and Cellular Biology.

[8]  B. O’Malley,et al.  Molecular mechanisms of action of steroid/thyroid receptor superfamily members. , 1994, Annual review of biochemistry.

[9]  Xing Zhang,et al.  Structure of the predominant protein arginine methyltransferase PRMT1 and analysis of its binding to substrate peptides. , 2003, Structure.

[10]  Yunbo Shi,et al.  Coactivator Recruitment Is Essential for Liganded Thyroid Hormone Receptor To Initiate Amphibian Metamorphosis , 2005, Molecular and Cellular Biology.

[11]  D. Aswad,et al.  Regulation of transcription by a protein methyltransferase. , 1999, Science.

[12]  Yunbo Shi,et al.  Transgenic Analysis Reveals that Thyroid Hormone Receptor Is Sufficient To Mediate the Thyroid Hormone Signal in Frog Metamorphosis , 2004, Molecular and Cellular Biology.

[13]  Y. B. Shi,et al.  Biphasic intestinal development in amphibians: embryogenesis and remodeling during metamorphosis. , 1996, Current topics in developmental biology.

[14]  C. Allis,et al.  Involvement of Histone Methylation and Phosphorylation in Regulation of Transcription by Thyroid Hormone Receptor , 2002, Molecular and Cellular Biology.

[15]  J. Wong,et al.  Coordinated Regulation of and Transcriptional Activation by Xenopus Thyroid Hormone and Retinoid X Receptors (*) , 1995, The Journal of Biological Chemistry.

[16]  Xiaodong Cheng,et al.  Crystal structure of the conserved core of protein arginine methyltransferase PRMT3 , 2000, The EMBO journal.

[17]  Tony Kouzarides,et al.  Crosstalk between CARM1 Methylation and CBP Acetylation on Histone H3 , 2002, Current Biology.

[18]  K. Yoshizato,et al.  Thyroid hormone regulation of a transcriptional coactivator in Xenopus laevis: Implication for a role in postembryonic tissue remodeling , 2002, Developmental dynamics : an official publication of the American Association of Anatomists.

[19]  Yunbo Shi,et al.  Distinct expression profiles of transcriptional coactivators for thyroid hormone receptors during Xenopus laevis metamorphosis , 2003, Cell Research.

[20]  M. Rosenfeld,et al.  Biological roles and mechanistic actions of co-repressor complexes. , 2002, Journal of cell science.

[21]  N. Koibuchi,et al.  TRAM-1, A Novel 160-kDa Thyroid Hormone Receptor Activator Molecule, Exhibits Distinct Properties from Steroid Receptor Coactivator-1* , 1997, The Journal of Biological Chemistry.

[22]  R. Evans,et al.  The steroid and thyroid hormone receptor superfamily. , 1988, Science.

[23]  Marc Montminy,et al.  A Transcriptional Switch Mediated by Cofactor Methylation , 2001, Science.

[24]  M. Stallcup,et al.  Synergistic Enhancement of Nuclear Receptor Function by p160 Coactivators and Two Coactivators with Protein Methyltransferase Activities* , 2001, The Journal of Biological Chemistry.

[25]  Mark T Bedford,et al.  Arginine methyltransferase CARM1 is a promoter‐specific regulator of NF‐κB‐dependent gene expression , 2005, The EMBO journal.

[26]  J. Wong,et al.  Transcriptional repression of Xenopus TR beta gene is mediated by a thyroid hormone response element located near the start site. , 1994, The Journal of biological chemistry.

[27]  Shih-Ming Huang,et al.  Synergistic, p160 Coactivator-dependent Enhancement of Estrogen Receptor Function by CARM1 and p300* , 2000, The Journal of Biological Chemistry.

[28]  Yunbo Shi,et al.  Tissue- and Gene-specific Recruitment of Steroid Receptor Coactivator-3 by Thyroid Hormone Receptor during Development*♦ , 2005, Journal of Biological Chemistry.

[29]  R. Roeder,et al.  The TRAP/SMCC/Mediator complex and thyroid hormone receptor function , 2001, Trends in Endocrinology & Metabolism.

[30]  T. Kouzarides,et al.  Methylation at arginine 17 of histone H3 is linked to gene activation , 2002, EMBO reports.

[31]  H. Matsuda,et al.  A Causative Role of Stromelysin-3 in Extracellular Matrix Remodeling and Epithelial Apoptosis during Intestinal Metamorphosis in Xenopus laevis* , 2005, Journal of Biological Chemistry.

[32]  P. Yen,et al.  Physiological and molecular basis of thyroid hormone action. , 2001, Physiological reviews.

[33]  Maho Takahashi,et al.  Coactivator-associated Arginine Methyltransferase 1, CARM1, Affects Pre-mRNA Splicing in an Isoform-specific Manner*♦ , 2005, Journal of Biological Chemistry.

[34]  Yunbo Shi,et al.  Molecular and developmental analyses of thyroid hormone receptor function in Xenopus laevis, the African clawed frog. , 2006, General and comparative endocrinology.

[35]  R. Koenig,et al.  A method for efficient production of recombinant thyroid hormone receptors reveals that receptor homodimer-DNA binding is enhanced by the coactivator TIF2. , 2005, Protein expression and purification.

[36]  M. Lazar Thyroid hormone receptors: multiple forms, multiple possibilities. , 1993, Endocrine reviews.

[37]  M. Y. Kim,et al.  Acetylation of estrogen receptor alpha by p300 at lysines 266 and 268 enhances the deoxyribonucleic acid binding and transactivation activities of the receptor. , 2006, Molecular endocrinology.

[38]  H. Gronemeyer,et al.  The coactivator TIF2 contains three nuclear receptor‐binding motifs and mediates transactivation through CBP binding‐dependent and ‐independent pathways , 1998, The EMBO journal.

[39]  N. Wong,et al.  Advances in our understanding of thyroid hormone action at the cellular level. , 1987, Endocrine reviews.

[40]  Hui Li,et al.  RAC3, a steroid/nuclear receptor-associated coactivator that is related to SRC-1 and TIF2. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[41]  R. Evans,et al.  Nuclear Receptor Coactivator ACTR Is a Novel Histone Acetyltransferase and Forms a Multimeric Activation Complex with P/CAF and CBP/p300 , 1997, Cell.

[42]  B. O’Malley,et al.  Sequence and Characterization of a Coactivator for the Steroid Hormone Receptor Superfamily , 1995, Science.

[43]  Wei Xu,et al.  Mutual synergistic folding in recruitment of CBP/p300 by p160 nuclear receptor coactivators , 2002, Nature.

[44]  H. Matsuda,et al.  Spatial and temporal expression profiles suggest the involvement of gelatinase A and membrane type 1 matrix metalloproteinase in amphibian metamorphosis , 2006, Cell and Tissue Research.

[45]  J. Wong,et al.  A role for cofactor–cofactor and cofactor–histone interactions in targeting p300, SWI/SNF and Mediator for transcription , 2003, The EMBO journal.

[46]  D. Trouche,et al.  Control of CBP co‐activating activity by arginine methylation , 2002, The EMBO journal.

[47]  Xiaodong Cheng,et al.  Synergy among Nuclear Receptor Coactivators: Selective Requirement for Protein Methyltransferase and Acetyltransferase Activities , 2002, Molecular and Cellular Biology.

[48]  Yunbo Shi,et al.  Recruitment of N-CoR/SMRT-TBLR1 Corepressor Complex by Unliganded Thyroid Hormone Receptor for Gene Repression during Frog Development , 2004, Molecular and Cellular Biology.

[49]  P. Jones,et al.  N-CoR-HDAC corepressor complexes: roles in transcriptional regulation by nuclear hormone receptors. , 2003, Current topics in microbiology and immunology.

[50]  G. Greene,et al.  Estrogen receptor accessory proteins: effects on receptor-DNA interactions. , 1995, Environmental health perspectives.

[51]  P. Chambon,et al.  TIF2, a 160 kDa transcriptional mediator for the ligand‐dependent activation function AF‐2 of nuclear receptors. , 1996, The EMBO journal.

[52]  Y. B. Shi,et al.  Cloning and characterization of the ribosomal protein L8 gene from Xenopus laevis. , 1994, Biochimica et biophysica acta.

[53]  M. Stallcup,et al.  Requirement for Multiple Domains of the Protein Arginine Methyltransferase CARM1 in Its Transcriptional Coactivator Function* , 2002, The Journal of Biological Chemistry.

[54]  H. Samuels,et al.  Regulation of gene expression by thyroid hormone. , 1988, The Journal of clinical investigation.

[55]  L. Freedman,et al.  Mediator complexes and transcription. , 2001, Current opinion in cell biology.

[56]  C. Lyttle,et al.  A Transcriptional Coactivator, Steroid Receptor Coactivator-3, Selectively Augments Steroid Receptor Transcriptional Activity* , 1998, The Journal of Biological Chemistry.

[57]  Yunbo Shi,et al.  Fusion Protein of Retinoic Acid Receptor α with Promyelocytic Leukemia Protein or Promyelocytic Leukemia Zinc Finger Protein Recruits N-CoR-TBLR1 Corepressor Complex to Repress Transcription in Vivo* , 2003, Journal of Biological Chemistry.

[58]  P. Meltzer,et al.  AIB1, a steroid receptor coactivator amplified in breast and ovarian cancer. , 1997, Science.

[59]  L. Freedman,et al.  Mechanisms of gene regulation by vitamin D(3) receptor: a network of coactivator interactions. , 2000, Gene.

[60]  M. Stallcup,et al.  GRIP1, a novel mouse protein that serves as a transcriptional coactivator in yeast for the hormone binding domains of steroid receptors. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[61]  K. Umesono,et al.  The nuclear receptor superfamily: The second decade , 1995, Cell.

[62]  Christopher K. Glass,et al.  The transcriptional co-activator p/CIP binds CBP and mediates nuclear-receptor function , 1997, Nature.

[63]  M.H.I. Dodd,et al.  10 – THE BIOLOGY OF METAMORPHOSIS , 1976 .