Advertisement

Modulation of Pre-mRNA Splicing Patterns with Synthetic Chemicals and Their Clinical Applications

  • Masatoshi Hagiwara
Conference paper

Abstract

Recent whole genome sequence analyses revealed that a high degree of proteomic complexity is achieved with a limited number of genes. This surprising finding underscores the importance of alternative splicing through which a single gene can generate structurally and functionally distinct protein isoforms [1]. Based on genome-wide analysis, 75% of human genes are thought to encode at least two alternatively spliced isoforms [2, 3]. The regulation of splice site usage provides a versatile mechanism for controlling gene expression and for the generation of proteome diversity, playing essential roles in many biological processes, such as embryonic development, cell growth, and apoptosis. The splice sites are generally recognized by the splicing machinery, a ribonuclear protein complex known as the spliceosome. Spliceosome binding is determined by competing activities of various auxiliary regulatory proteins, such as members of SR protein or heterogeneous nuclear ribonucleoprotein (hnRNP) protein families, which bind specific regulatory sequences and alter the binding of the spliceosome to a particular splice site [1, 4]. Pre-mRNA splicing is regulated in a tissue-specific or developmental stage-specific manner [5]. The selection of splice site can be altered by numerous extracellular stimuli such as hormones, immune response, neuronal depolarization, and cellular stress, through changes in synthesis/degradation, complex formation, and intracellular localization of regulatory proteins. SR proteins are heavily phosphorylated in cells and involved in constitutive and alternative splicing, and the phosphorylation states of SR proteins are altered in response to these extracellular stimuli [6]. Splicing mutations located in either intronic or exonic regions frequently cause hereditary diseases, and more than 15% of mutations that cause genetic disease affect pre-mRNA splicing [7]. Based on a hypothetical idea that we can cure human diseases by regulating the phosphorylation state of SR proteins with synthetic inhibitors of protein kinases, we started our long voyage to challenge the development of new chemical therapeutics.

Keywords

Splice Site Down Syndrome Duchenne Muscular Dystrophy Severe Acute Respiratory Syndrome Duchenne Muscular Dystrophy 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

The author thanks past and present Hagiwara’s laboratory members and collaborators, especially T. Hosoya, A. Nishida, D.G. Nowak, D.O. Bates, M. Matsuo, A. Anwar, M.A. Garcia-Blanco, T. Fukuhara, M. Muraki, N. Kataoka, Y. Ogawa, and H. Onogi, for their contribution to the work described here. This work was supported by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) of Japan and research grants from the Japan Science and Technology Agency, Takeda Science Foundation, The Naito Foundation Natural Science Scholarship, and The Uehara Memorial Foundation.

References

  1. 1.
    Black DL (2003) Mechanisms of alternative pre-messenger RNA splicing. Annu Rev Biochem 72:291–336PubMedCrossRefGoogle Scholar
  2. 2.
    Modrek B, Lee CJ (2003) Alternative splicing in the human, mouse and rat genomes is associated with an increased frequency of exon creation and/or loss. Nat Genet 34:177–180PubMedCrossRefGoogle Scholar
  3. 3.
    Johnson JM, Castle J, Garrett-Engele P et al (2003) Genome-wide survey of human alternative pre-mRNA splicing with exon junction microarrays. Science 302:2141–2144PubMedCrossRefGoogle Scholar
  4. 4.
    Smith CW, Valcarcel J (2000) Alternative pre-mRNA splicing: the logic of combinatorial control. Trends Biochem Sci 25:381–388PubMedCrossRefGoogle Scholar
  5. 5.
    Stamm S (2002) Signals and their transduction pathways regulating alternative splicing: a new dimension of the human genome. Hum Mol Genet 11:2409–2416PubMedCrossRefGoogle Scholar
  6. 6.
    Long JC, Caceres JF (2009) The SR protein family of splicing factors: master regulators of gene expression. Biochem J 417:15–27PubMedCrossRefGoogle Scholar
  7. 7.
    Krawczak M, Reiss J, Cooper DN (1992) The mutational spectrum of single base-pair substitutions in mRNA splice junctions of human genes: causes and consequences. Hum Genet 90:41–54PubMedCrossRefGoogle Scholar
  8. 8.
    Fukuhara T, Hosoya T, Shimizu S, Sumi K, Oshiro T, Yoshinaka Y, Suzuki M, Yamamoto N, Herzenberg LA, Herzenberg LA, Hagiwara M (2006) Utilization of host SR protein kinases and RNA-splicing machinery during viral replication. Proc Natl Acad Sci USA 103:11329–11333PubMedCrossRefGoogle Scholar
  9. 9.
    Muraki M, Ohkawara B, Hosoya T, Onogi H, Koizumi J, Koizumi T, Sumi K, Yomoda J, Murray MV, Kimura K, Furuichi K, Shibuya H, Krainer AR, Suzuki M, Hagiwara M (2004) Manipulation of alternative splicing by a newly developed inhibitor of Clks. J Biol Chem 279:24246–24254PubMedCrossRefGoogle Scholar
  10. 10.
    Anwar A, Hosoya T, Leong KM, Onogi H, Okuno Y, Hiramatsu T, Koyama H, Suzuki M, Hagiwara M, Garcia-Blanco MA (2011) The Kinase Inhibitor SFV785 Dislocates Dengue Virus Envelope Protein from the Repli­cation Complex and Blocks Virus Assembly. PLoS One 6(8):e23246CrossRefGoogle Scholar
  11. 11.
    Karlas A, Machuy N, Shin Y, Pleissner KP, Artarini A, Heuer D, Becker D, Khalil H, Ogilvie LA, Hess S, Mäurer AP, Müller E, Wolff T, Rudel T, Meyer TF (2010) Genome-wide RNAi screen identifies human host factors crucial for influenza virus replication. Nature (Lond) 463:818–822CrossRefGoogle Scholar
  12. 12.
    Harper SJ, Bates DO (2008) VEGF-A splicing: the key to anti-angiogenic therapeutics? Nat Rev Cancer 8:880–887PubMedCrossRefGoogle Scholar
  13. 13.
    Nowak DG, Amin EM, Rennel ES, Hoareau-Aveilla C, Gammons M, Damodoran G, Hagiwara M, Harper SJ, Woolard J, Ladomery MR, Bates DO (2010) Regulation of vascular endothelial growth factor (VEGF) splicing from pro-angiogenic to anti-angiogenic isoforms: a novel therapeutic strategy for angiogenesis. J Biol Chem 285:5532–5540PubMedCrossRefGoogle Scholar
  14. 14.
    Nishida A, Kataoka N, Takeshima Y, Yagi M, Awano H, Ota M, Itoh K, Hagiwara M, Matsuo M (2011) Chemical treatment enhances skipping of a mutated exon in the dystrophin gene. Nat Commun 2:308PubMedCrossRefGoogle Scholar
  15. 15.
    Duncan PI, Howell BW, Marius RM, Drmanic S, Douville EM, Bell JC (1995) Alternative splicing of STY, a nuclear dual specificity kinase. J Biol Chem 270:21524–21531PubMedCrossRefGoogle Scholar
  16. 16.
    Ninomiya K, Kataoka N, Hagiwara M (2011) Stress-responsive maturation of Clk1/4 pre-mRNAs promotes phosphorylation of SR splicing factor. J Cell Biol 195:27–40PubMedCrossRefGoogle Scholar
  17. 17.
    Wiseman FK, Alford KA, Tybulewicz VL, Fisher EM (2009) Down syndrome: recent progress and future prospects. Hum Mol Genet 18:R75–R83PubMedCrossRefGoogle Scholar
  18. 18.
    Dowjat WK, Adayev T, Kuchna I et al (2007) Trisomy-driven overexpression of DYRK1A kinase in the brain of subjects with Down syndrome. Neurosci Lett 413:77–81PubMedCrossRefGoogle Scholar
  19. 19.
    Ogawa Y, Nonaka Y, Goto T, Ohnishi E, Hiramatsu T, Kii I, Yoshida M, Ikura T, Onogi H, Shibuya H, Hosoya T, Ito N, Hagiwara M (2010) Development of a novel selective inhibitor of the Down syndrome-related kinase Dyrk1A. Nat Commun 1:86PubMedCrossRefGoogle Scholar

Copyright information

© Springer 2012

Authors and Affiliations

  1. 1.Department of Anatomy and Developmental Biology, Graduate School of MedicineKyoto UniversityKyotoJapan

Personalised recommendations