1. Some references of interest on Photogenetics 1)Structure and Function of Plant Photoreceptors. Möglich, Yang, Ayers, Moffat. Annual Review of Plant Biology, online (in press) 2010. 2)Design and signaling mechanisms of light regulated histidine kinases. Möglich, Ayers, Moffat. J. Mol Biol., 385: 1433, 2009. 3)Generation of new protein functions by nonhomologous combinations and rearrangements of domains and modules. Shohei. Current Opinion in Biotechnology, 20:398. 2009. 4)Genetically encoded photoswitching of actin assembly through the Cdc42-WAS-ARP2/3 complex pathway. Leung, Otomo, Chory, Rosen. PNAS, 105, 12797, 2008. 5)Light activated DNA binding in a designed allosteric protein. Strickland, Moffat, Sosnick. PNAS 105:10709, 2008.

2. Structure and Function of Plant Photoreceptors 2010 Annual Review by Möglich, Yang, Ayers, Moffat Structural changes in A. sativa Phototropin1 Lov2 domain in response to blue light. Not shown : NMR data indicate that the Jα C terminal helix unfolds when the thioether bound is formed and there are changes in hydrogen bounding.

3. If the Lov domain polypeptide covers a binding site on a protein and such that protein-protein interaction is impaired in the dark, then in the response to low fluence blue light, the Jα domain should unfold and flip the Lov domain out of the way to reveal the binding site.

4. Dark Light

5. Connecting Leu 546 of Lov2-Jα to Ile4 of Rac1 reduces Rac binding to PAK Tested PAK (effector) binding to Photoactivated Rac1 (PA-Rac1) Lov2-Jα (AA 407-547) was fused to N-terminus of Rac1 Light insensitive “lit” state mutant

8. YI Wu et al. Nature 000, 1-5 (2009) doi:10.1038/nature08241 Crystallization and structural modelling of PA-Rac1. Lov2 JαJα Rac1 Photoactive(PA)-Cdc42 binds PAK “Lit” state No inhibition Trp mutation decreases PAK binding Adding the PAK CRIB domain inhibits binding

9. Photogenetics can be used to light or dark activate. N 2 fixing bacteria oxygen-sensitive kinase reprogramed to be active in the dark by replacing the oxygen sensor in the kinase with LOV domain. Light reduced kinase activity 1000 fold in vitro. Rational design for photo-sensing proteins. Design and signaling mechanisms of light regulated histidine kinases. Möglich, Ayers, Moffat. J. Mol Biol., 385: 1433, 2009.

10. Structure and Function of Plant Photoreceptors 2010 Annual Review by Möglich, Yang, Ayers, Moffat

11. What types of chemical or photosensors can you develop for your research?

14. Engineering a genetically encoded, reversible light-controlled signaling system Leung D W et al. PNAS 2008;105:12797-12802 ©2008 by National Academy of Sciences

15. YI Wu et al. Nature 000, 1-5 (2009) doi:10.1038/nature08241 Localized activation or inactivation of PA-Rac1 induces myosin-dependent migration.

16. YI Wu et al. Nature 000, 1-5 (2009) doi:10.1038/nature08241 Inhibition of RhoA by PA-Rac1.