1. CZ.1.07/2.3.00/30.0037 Employment of Best Young Scientists for International Cooperation Empowerment 2 nd Postdoc Presentation Day 14. 3. 2014 Name of the postdoc: Peter Kolesár

2. Project title: Ubiquitin ligase activity of the Smc5/6 complex Mentor: doc. Mgr. Jan Paleček, Dr. rer. nat. Faculty/Department/Research group: CEITEC / Genomics and Proteomics of Plant Systems / Chromatin Molecular Complexes Start date of postdoc (in the project): 09/2013 2

3. Smc5/6 complex 3 White & Erickson, JSB (2006) SMC6SMC5 Nse3 Nse1

4. 4 In vitro ubiquitination assay (human) Woels et al., 2007

5. hNse1 is autoubiquitnated (K143, K144) 5 hNse3 is ubiquitnated at K83, K220, K221

6. Nse1 ubiquitin ligase activity is conserved - S. pombe enzymes: 6

7. SpNse3 is ubiquitinated 7

8. Nse1 without RING loses its E3 activity (S. pombe) 8

9. 9 PLANS Determine the ubiquitinated targets of Nse1 – in vitro assay (Nse1,2,3,4, Smc5,6, Brc1, Mis4,H4,..) - Nse1-TAP tag purification - SILAC – Nse1wt vs. Nse1-RING* Determine the consequences of Nse1 E3 activity - mutate modified lysines and determine phenotypes

10. How and where are you going to publish your results (1 slide) 10