MECHANISM OF NUCLEOSOME DISASSEMBLY BY HISTONE CHAPERONES

PhD student project 1
(Early Stage Researcher 1, ESR1)

MECHANISM OF NUCLEOSOME DISASSEMBLY BY HISTONE CHAPERONES

Supervision: Rolf Boelens and Hugo van Ingen

Early Stage Researcher: Ivan Corbeski

       

Aim:
Uncover the mechanism of nucleosome assembly and disassembly.

Methodology:
ESR1 learns to use in vitro reconstituted nucleosomes and chaperones and advanced MeTROSY NMR techniques, to answer these questions: (i) What are the kinetics of histone eviction and are these related to intrinsic DNA unwrapping? (ii) What are changes in DNA dynamics concomitant with histone eviction? (iii) What changes occur in structure and dynamics of the remaining DNA-bound histones once part of the histone are evicted? (iv) Can we find experimental evidence for the proposed ‘strand-capture’ model for ASF1 mediated H3-H4 eviction? (v) What are parallels between salt-dependent disassembly and chaperone mediated disassembly? (Collaborations Heck, Laue, SpronkNMR)

Project goals:
Production, biophysical and NMR characterization of a suitable nucleosomal chaperone (NC). NMR interaction studies of NC and nucleosome. Biochemical and biophysical (including NMR) studies of nucleosome assembly/disassembly.

Collaborators:
Heck; Laue (Cambridge); SpronkNMR

Key publications:

  1. R.G. Hibbert, A. Huang, R. Boelens* and T.K. Sixma* (2011) # joint corresponding authors ‘E3 ligase Rad18 promotes monoubiquitination rather than ubiquitin chain formation by E2 enzyme Rad6’, Proc Natl Acad Sci USA 108, 5590-5595
  2. M. Sette, R. Spurio, E. Trotta, A. Brandi, C.L. Pon, G. Barbato, R. Boelens* and C.O. Gualerzi* (2009) # joint corresponding authors ‘DNA-binding specificity of the C-terminal domain of nucleoid-associated protein H-NS’, J Biol Chem 284, 30453-30462.
  3. H. van Ingen, F.M.A. van Schaik, H. Wienk, J. Ballering, H. Rehmann, A.C. Dechesne, J.A.W. Kruijzer, R.M.J. Liskamp, H.Th.M. Timmers, R. Boelens (2008), ‘Structural Insight into the Recognition of the H3K4me3 mark by the TFIID subunit TAF3’, Structure 16 , 1245-1256.