Role of methyl-cytosine binding proteins in the gain and loss of pluripotency

• Applicant: Professorin Dr. Maria Cristina Cardoso
• Subject Area: Developmental Biology ; Cell Biology
• Term: from 2008 to 2015
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 66434331

• Final Report Year: 2014

Final Report Abstract

During the course of this project and in collaboration with many colleagues within and outside the priority program, we generated a series of biological materials and tools, as well as novel methodological approaches and new computational protocols. With these tools we systematically investigated MeCP2 level and distribution in totipotent, pluripotent, differentiated and reprogrammed (iPS) cells and in tissues. We examined the binding and dynamics of wild type and mutant forms of MeCP2 at chromatin and tested whether mutants of MeCP2 could be rescued by targeting ectopically to chromatin. In addition, we investigated whether MBDs bind to newly reported cytosine modifications and how the level of MBDs and their interactions influences chromatin higher order organization and gene activity. The development of new computational approaches allowed us to study the correlation of spatial nuclear (re)positioning of genes and their expression level during gain and loss of pluripotency. Altogether the biological, methodological and computational tools we developed were and will be very important to investigate and understand the regulation of methyl CpG binding domain proteins but also, on a broader level, the functional nuclear and genome architecture. In summary, our results contribute to our understanding of the role of MBDs in shaping higher order genome organization and dynamics in pluripotency and reprogramming.

Publications

• (2011). 3D-Image analysis platform monitoring relocation of pluripotency genes during reprogramming. Nucleic Acids Res. 39: e113
  Jost, K. L., Haase, S., Smeets, D., Schrode, N., Schmiedel, J. M., Bertulat, B., Hertel, H., Cremer, M. and Cardoso, M. C.
  (See online at https://doi.org/10.1093/nar/gkr486)
• (2011). Generation and characterization of rat and mouse monoclonal antibodies specific for MeCP2 and their use in X-inactivation studies. PLoS ONE 6: e26499
  Jost, K. L., Rottach, A., Milden, M., Bertulat, B., Becker, A., Wolf, P., Sandoval, J., Petazzi, P., Huertas, D., Esteller, M., Kremmer, E., Leonhardt, H. and Cardoso, M. C.
  (See online at https://doi.org/10.1371/journal.pone.0026499)
• (2011). MeCP2 Rett mutations affect large scale chromatin organization. Hum. Mol. Genet. 20: 4187-4195
  Agarwal, N., Becker, A., Jost, K. L., Haase, S., Thakur, B. K., Brero, A., Hardt, T., Kudo, S., Leonhardt, H. and Cardoso, M. C.
  (See online at https://doi.org/10.1093/hmg/ddr346)
• (2011). Recognition of 5-hydroxymethylcytosine by the Uhrf1 SRA domain. PLoS ONE 6: e21306
  Frauer, C., Hoffmann, T. Bultmann, S., Casa, V., Cardoso, M. C., Antes, I. and Leonhardt, H.
  (See online at https://doi.org/10.1371/journal.pone.0021306)
• (2012). Heterochromatin and gene positioning: inside, outside, any side? Chromosoma 121: 555-563
  Jost, K. L., Bertulat, B. and Cardoso, M. C.
  (See online at https://doi.org/10.1007/s00412-012-0389-2)
• (2012). MeCP2 dependent heterochromatin reorganization during neural differentiation of a novel MeCP2-deficient embryonic stem cell reporter line. PLoS ONE 7: e47848
  Bertulat, B., De Bonis, M. L., Della Ragione, F., Lehmkuhl, A., Milden, M., Storm, C., Jost, K. L., Scala, S., Hendrich, B., D’Esposito, M. and Cardoso, M. C.
  (See online at https://doi.org/10.1371/journal.pone.0047848)
• (2012). Targeted manipulation of heterochromatin rescues MeCP2 Rett mutants and re-establishes higher-order chromatin organization. Nucleic Acids Res. 40: e176
  Casas-Delucchi, C. S., Becker, A, Bolius, J. J. and Cardoso, M. C.
  (See online at https://doi.org/10.1093/nar/gks784)
• (2013). A role for MeCP2 in switching gene activity via chromatin unfolding and HP1γ displacement. PLoS ONE 8: e69347
  Brink, M. C., Piebes, D. G. E., de Groote, M. L., Luijsterburg, M. S., Casas-Delucchi, C. S., van Driel, R., Rots, M. G., Cardoso, M. C. and Verschure, P. J.
  (See online at https://doi.org/10.1371/journal.pone.0069347)
• (2013). Direct homo- and hetero-interactions of MeCP2 and MBD2. PLoS ONE 8: e53730
  Becker, A., Allmann, L., Hofstätter, M., Casà, V., Weber, P., Lehmkuhl, A., Herce, H. D. and Cardoso, M. C.
  (See online at https://doi.org/10.1371/journal.pone.0053730)
• (2014). DNA methylation reader MECP2: cell type- and differentiation stage-specific distribution. Epigenetics Chromatin 7: 17
  Song, C., Feodorova, Y., Guy, J., Peichl, L., Jost, K. L., Kimura, H., Cardoso, M. C., Bird, A., Leonhardt, H., Joffe, B. and Solovei, I.
  (See online at https://doi.org/10.1186/1756-8935-7-17)