University of Manchester, Faculty of Medicine, and Health Sciences 3-Year Post-doctoral position in biology “Quantification of dynamics driving the ” Although circadian biology has been driven by a revolution in genetics we have generally been unable to study the encoded that actually make up the core circadian clockwork in cells: we still do not understand how they behave in circadian time, or across cellular space. Building on our earlier success with mammalian PER2 protein (Smyllie et al 2016), we will now simultaneously track dynamic changes in multiple circadian clock proteins in real-time in single cells in brain (SCN neurons) and peripheral cells. The project combines state-of-the-art CRISPR editing to tag multiple clock proteins with different fluorophores, real-time single cell imaging and spectroscopy to quantify dynamic changes in protein interactions and concentration. We will study the effects of mutations that alter circadian pace-making, and explore how pharmacological manipulation drives the behaviour of clock complexes in peripheral and brain (SCN) clock neurons. With these tools, we aim to obtain mechanistic understanding and generate new models with predictive power in this complex dynamic system, likely serving as an exemplar for quantitative well beyond the circadian domain. The project combines the laboratories of Michael Hastings (MRC Laboratory of , Cambridge), Mike White, Dave Spiller and Andrew Loudon (Manchester), plus collaborations with Carrie Partch (University of California, Santa Cruz) and Carla Green (UT Southwestern). This post is based at Manchester, and the successful applicant will be expected to work closely with a post-doctoral scientist based at Cambridge and also colleagues in the USA. We seek to appoint between October and December 2017. For further details, contact Prof Andrew [email protected]; Key webs sites: https://www.bmh.manchester.ac.uk/research/biological-timing/ http://www2.mrc-lmb.cam.ac.uk/group-leaders/h-to-m/michael-hastings/

Key recent papers:

Brancaccio M, Patton AP, Chesham JE, Maywood ES, Hastings MH. (2017) Astrocytes Control Circadian Timekeeping in the via Glutamatergic Signaling. Neuron;93(6):1420-1435

Smyllie NJ, Pilorz V, Boyd J, Meng QJ, Saer B, Chesham JE, Maywood ES, Krogager TP, Spiller DG, Boot- Handford R, White MR, Hastings MH, Loudon AS (2016) Visualizing and Quantifying Intracellular Behavior and Abundance of the Core Circadian Clock Protein PERIOD2. Curr Biol;26(14):1880-6.

Smyllie NJ, Chesham JE, Hamnett R, Maywood ES, Hastings MH. (2016) Temporally chimeric mice reveal flexibility of circadian period-setting in the suprachiasmatic nucleus. Proc Natl Acad Sci U S A. 113(13):3657- 62.

Adamson A, Boddington C, Downton P, Rowe W, Bagnall J, Lam C, Maya-Mendoza A, Schmidt L, Harper CV, Spiller DG, Rand DA, Jackson DA, White MR, Paszek P. Signal transduction controls heterogeneous NF-κB dynamics and target through cytokine-specific refractory states. Nat Commun. 2016 Jul 6;7:12057