DYNAMIC PROFILING OF THE KAIABC CIRCADIAN CLOCK SYSTEM

PhD student project 6
(Early Stage Researcher 6, ESR6)

DYNAMIC PROFILING OF THE KAIABC CIRCADIAN CLOCK SYSTEM

Supervision: Albert Heck

Early Stage Researcher: Philip Lössl
 

   

Aim:
Understanding how dynamic protein assembly regulates the cyanobacterial clock.

Methodology:
ESR6 will learn to unravel features of the cyanobacterial circadian clock at the molecular level, taking complementary mass spectrometry based approaches focusing on novel structural aspects of the KaiABC system. Using native (ion mobility) mass spectrometry, HD exchange and cross-linking mass spectrometry analyses binary and ternary complexes formed by the KaiABC system, and how all co-existing protein complexes are regulated during the circadian rhythm (collaboration MSVision). ESR6 will be able to monitor in parallel co-occurring protein assemblies and the dynamic exchange of subunits, but also determine cross-sections of the complexes and their sub-complexes. Using this data, combined with computational modelling and EM, ESR6 will produce new structural models and provide data on how these distinct protein complexes may play a role in the circadian phosphorylation cycle of the KaiABC system (collaboration Gros, Thomas). ESR6 will also investigate a much less studied even more minimal circadian clock that lacks the KaiA gene.

Collaborators:
Gros; Thomas (McGill); MSVision

Project goals:
New insights into the structural biology of the Kai system and how that relates to the system-wide regulation of the circadian rhythm.

Key publications:

  1. C. Uetrecht, R.J. Rose, E. van Duijn, K. Lorenzen and A.J.R. Heck (2010) ‘Ion mobility mass spectrometry of proteins and protein assemblies.’ Chem Soc Rev 39, 1633-55.
  2. G.K. Shoemaker, E. van Duijn, S.E. Crawford, C. Uetrecht, M. Baclayon, W.H. Roos, G.J. Wuite, M.K. Estes, B.V. Prasad and A.J.R. Heck (2010) ‘Norwalk virus assembly and stability monitored by mass spectrometry.’ Mol Cell Proteomics 9, 1742-51.
  3. C. Uetrecht, C. Versluis, N.R. Watts, P.T. Wingfield, A.C. Steven and A.J.R. Heck (2008) ‘Stability and shape of hepatitis B virus capsids in vacuo.’ Angew Chem Int Ed Engl 47, 6247-51.