Advertisement

Journal of Molecular Neuroscience

, Volume 51, Issue 2, pp 615–628 | Cite as

Alternative Splicing of the Chromodomain Protein Morf4l1 Pre-mRNA Has Implications on Cell Differentiation in the Developing Chicken Retina

  • Henrik Boije
  • Henrik Ring
  • Shahrzad Shirazi Fard
  • Ida Grundberg
  • Mats Nilsson
  • Finn Hallböök
Article

Abstract

The proliferation, cell cycle exit and differentiation of progenitor cells are controlled by several different factors. The chromodomain protein mortality factor 4-like 1 (Morf4l1) has been ascribed a role in both proliferation and differentiation. Little attention has been given to the existence of alternative splice variants of the Morf4l1 mRNA, which encode two Morf41l isoforms: a short isoform (S-Morf4l1) with an intact chromodomain and a long isoform (L-Morf4l1) with an insertion in or in the vicinity of the chromodomain. The aim of this study was to investigate if this alternative splicing has a function during development. We analysed the temporal and spatial distribution of the two mRNAs and over-expressed both isoforms in the developing retina. The results showed that the S-Morf4l1 mRNA is developmentally regulated. Over-expression of S-Morf4l1 using a retrovirus vector produced a clear phenotype with an increase of early-born neurons: retinal ganglion cells, horizontal cells and cone photoreceptor cells. Over-expression of L-Morf4l1 did not produce any distinguishable phenotype. The over-expression of S-Morf4l1 but not L-Morf4l1 also increased apoptosis in the infected regions. Our results suggest that the two Morf4l1 isoforms have different functions during retinogenesis and that Morf4l1 functions are fine-tuned by developmentally regulated alternative splicing. The data also suggest that Morf4l1 contributes to the regulation of cell genesis in the retina.

Keywords

Acetylation Avian Chromatin structure Development HAT HDAC Isoform Histon MRG15 MRGX Neuron RCAS Retina Splicing Virus vector 

Notes

Acknowledgments

We thank Karl Wahlin for the gateway-adapted RCAS vector, Pernilla Bjerling for the discussions and input. The work was supported by the Swedish Research Council (20859-01-3, 12187-15-3), Barncancerfonden (PROJ09/038), Ögonfonden, St Eriks ögonsjukhus stipendier and Kronprinsessan Margaretas arbetsnämnd för synskadade.

References

  1. Akhtar A, Becker PB (2000) Activation of transcription through histone H4 acetylation by MOF, an acetyltransferase essential for dosage compensation in Drosophila. Mol Cell 5:367–375PubMedCrossRefGoogle Scholar
  2. Akhtar A, Zink D, Becker PB (2000) Chromodomains are protein-RNA interaction modules. Nature 407:405–409PubMedCrossRefGoogle Scholar
  3. Baner J, Nilsson M, Mendel-Hartvig M, Landegren U (1998) Signal amplification of padlock probes by rolling circle replication. Nucleic Acids Res 26:5073–5078PubMedCrossRefGoogle Scholar
  4. Bertram MJ, Pereira-Smith OM (2001) Conservation of the MORF4 related gene family: identification of a new chromo domain subfamily and novel protein motif. Gene 266:111–121PubMedCrossRefGoogle Scholar
  5. Bertram MJ, Berube NG, Hang-Swanson X et al (1999) Identification of a gene that reverses the immortal phenotype of a subset of cells and is a member of a novel family of transcription factor-like genes. Mol Cell Biol 19:1479–1485PubMedGoogle Scholar
  6. Boardman PE, Sanz-Ezquerro J, Overton IM et al (2002) A comprehensive collection of chicken cDNAs. Curr Biol 12:1965–1969PubMedCrossRefGoogle Scholar
  7. Boije H, Edqvist PH, Hallbook F (2008) Temporal and spatial expression of transcription factors FoxN4, Ptf1a, Prox1, Isl1 and Lim1 mRNA in the developing chick retina. Gene Expr Patterns 8:117–123PubMedCrossRefGoogle Scholar
  8. Boije H, Ring H, Lopez-Gallardo M, Prada C, Hallbook F (2010) Pax2 is expressed in a subpopulation of Muller cells in the central chick retina. Dev Dyn 239:1858–1866PubMedCrossRefGoogle Scholar
  9. Boije H, Shirazi Fard S, Ring H, Hallbook F (2013) Forkheadbox N4 (FoxN4) triggers context-dependent differentiation in the developing chick retina and neural tube. Differentiation 85:11–19PubMedCrossRefGoogle Scholar
  10. Castedo M, Perfettini JL, Roumier T, Andreau K, Medema R, Kroemer G (2004) Cell death by mitotic catastrophe: a molecular definition. Oncogene 23:2825–2837PubMedCrossRefGoogle Scholar
  11. Chen M, Takano-Maruyama M, Pereira-Smith OM, Gaufo GO, Tominaga K (2009) MRG15, a component of HAT and HDAC complexes, is essential for proliferation and differentiation of neural precursor cells. J Neurosci Res 87:1522–1531PubMedCrossRefGoogle Scholar
  12. Chen M, Tominaga K, Pereira-Smith OM (2010) Emerging role of the MORF/MRG gene family in various biological processes, including aging. Ann N Y Acad Sci 1197:134–141PubMedCrossRefGoogle Scholar
  13. Chen M, Pereira-Smith OM, Tominaga K (2011) Loss of the chromatin regulator MRG15 limits neural stem/progenitor cell proliferation via increased expression of the p21 Cdk inhibitor. Stem Cell Res 7:75–88PubMedCrossRefGoogle Scholar
  14. Edqvist PH, Lek M, Boije H, Lindback SM, Hallbook F (2008) Axon-bearing and axon-less horizontal cell subtypes are generated consecutively during chick retinal development from progenitors that are sensitive to follistatin. BMC Dev Biol 8:46PubMedCrossRefGoogle Scholar
  15. ENSGT00530000063018 (2012) MORF4 gene tree. http://www.ensembl.org/Multi/GeneTree?gt=ENSGT00530000063018. Accessed 14 June 2012
  16. Fisher CL, Fisher AG (2011) Chromatin states in pluripotent, differentiated, and reprogrammed cells. Curr Opin Genet Dev 21:140–146PubMedCrossRefGoogle Scholar
  17. Hamburger V, Hamilton HL (1992) A series of normal stages in the development of the chick embryo. 1951. Dev Dyn 195:231–272PubMedCrossRefGoogle Scholar
  18. HGNC (2012) MORF4. HUGO gene nomenclature committee at the European bioinformatics Institute. http://www.genenames.org/data/hgnc_data.php?hgnc_id=15773. Accessed 14 June 2012
  19. Ka S, Kerje S, Bornold L, Liljegren U, Siegel PB, Andersson L, Hallböök F (2009) Proviral integrations and expression of endogenous Avian leucosis virus during long term selection for high and low body weight in two chicken lines. Retrovirology 6:68PubMedCrossRefGoogle Scholar
  20. Kim D, Blus BJ, Chandra V, Huang P, Rastinejad F, Khorasanizadeh S (2010) Corecognition of DNA and a methylated histone tail by the MSL3 chromodomain. Nat Struct Mol Biol 17:1027–1029PubMedCrossRefGoogle Scholar
  21. Kranenburg O, van der Eb AJ, Zantema A (1996) Cyclin D1 is an essential mediator of apoptotic neuronal cell death. EMBO J 15:46–54PubMedGoogle Scholar
  22. Kumar GS, Chang W, Xie T et al (2012) Sequence requirements for combinatorial recognition of histone H3 by the MRG15 and Pf1 subunits of the Rpd3S/Sin3S corepressor complex. J Mol Biol 422:519–531PubMedCrossRefGoogle Scholar
  23. Larsson C, Grundberg I, Soderberg O, Nilsson M (2010) In situ detection and genotyping of individual mRNA molecules. Nat Methods 7:395–397PubMedCrossRefGoogle Scholar
  24. Lee EY, Chang CY, Hu N et al (1992) Mice deficient for Rb are nonviable and show defects in neurogenesis and haematopoiesis. Nature 359:288–294PubMedCrossRefGoogle Scholar
  25. Lessard J, Wu JI, Ranish JA et al (2007) An essential switch in subunit composition of a chromatin remodeling complex during neural development. Neuron 55:201–215PubMedCrossRefGoogle Scholar
  26. Leung JK, Berube N, Venable S, Ahmed S, Timchenko N, Pereira-Smith OM (2001) MRG15 activates the B-myb promoter through formation of a nuclear complex with the retinoblastoma protein and the novel protein PAM14. J Biol Chem 276:39171–39178PubMedCrossRefGoogle Scholar
  27. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T))-method. Methods 25:402–408PubMedCrossRefGoogle Scholar
  28. Loftus SK, Larson DM, Watkins-Chow D, Church DM, Pavan WJ (2001) Generation of RCAS vectors useful for functional genomic analyses. DNA Res 8:221–226PubMedCrossRefGoogle Scholar
  29. Matsuoka Y, Matsuoka Y, Shibata S et al (2002) A chromodomain-containing nuclear protein, MRG15 is expressed as a novel type of dendritic mRNA in neurons. Neurosci Res 42:299–308PubMedCrossRefGoogle Scholar
  30. Mey J, Thanos S (2000) Development of the visual system of the chick. I. Cell differentiation and histogenesis. Brain Res Brain Res Rev 32:343–379PubMedCrossRefGoogle Scholar
  31. Pardo PS, Leung JK, Lucchesi JC, Pereira-Smith OM (2002) MRG15, a novel chromodomain protein, is present in two distinct multiprotein complexes involved in transcriptional activation. J Biol Chem 277:50860–50866PubMedCrossRefGoogle Scholar
  32. Pena AN, Tominaga K, Pereira-Smith OM (2011) MRG15 activates the cdc2 promoter via histone acetylation in human cells. Exp Cell Res 317:1534–1540PubMedCrossRefGoogle Scholar
  33. Qin XQ, Livingston DM, Kaelin WG Jr, Adams PD (1994) Deregulated transcription factor E2F-1 expression leads to S-phase entry and p53-mediated apoptosis. Proc Natl Acad Sci U S A 91:10918–10922PubMedCrossRefGoogle Scholar
  34. Smith ER, Cayrou C, Huang R, Lane WS, Cote J, Lucchesi JC (2005) A human protein complex homologous to the Drosophila MSL complex is responsible for the majority of histone H4 acetylation at lysine 16. Mol Cell Biol 25:9175–9188PubMedCrossRefGoogle Scholar
  35. Tominaga K, Pereira-Smith OM (2002) The genomic organization, promoter position and expression profile of the mouse MRG15 gene. Gene 294:215–224PubMedCrossRefGoogle Scholar
  36. Tominaga K, Leung JK, Rookard P, Echigo J, Smith JR, Pereira-Smith OM (2003) MRGX is a novel transcriptional regulator that exhibits activation or repression of the B-myb promoter in a cell type-dependent manner. J Biol Chem 278:49618–49624PubMedCrossRefGoogle Scholar
  37. Tominaga K, Kirtane B, Jackson JG et al (2005a) MRG15 regulates embryonic development and cell proliferation. Mol Cell Biol 25:2924–2937PubMedCrossRefGoogle Scholar
  38. Tominaga K, Matzuk MM, Pereira-Smith OM (2005b) MrgX is not essential for cell growth and development in the mouse. Mol Cell Biol 25:4873–4880PubMedCrossRefGoogle Scholar
  39. Yochum GS, Ayer DE (2002) Role for the mortality factors MORF4, MRGX, and MRG15 in transcriptional repression via associations with Pf1, mSin3A, and Transducin-Like Enhancer of Split. Mol Cell Biol 22:7868–7876PubMedCrossRefGoogle Scholar
  40. Zhang P, Du J, Sun B et al (2006) Structure of human MRG15 chromo domain and its binding to Lys36-methylated histone H3. Nucleic Acids Res 34:6621–6628PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Henrik Boije
    • 1
  • Henrik Ring
    • 1
  • Shahrzad Shirazi Fard
    • 1
  • Ida Grundberg
    • 2
    • 3
  • Mats Nilsson
    • 2
    • 4
  • Finn Hallböök
    • 1
  1. 1.Department of Neuroscience, BMCUppsala UniversityUppsalaSweden
  2. 2.Department of Immunology, Genetics and Pathology, Rudbeck LaboratoryUppsala UniversityUppsalaSweden
  3. 3.Olink BioscienceUppsalaSweden
  4. 4.Science for Life Laboratory, Department of Biochemistry and BiophysicsStockholm UniversityStockholmSweden

Personalised recommendations