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.

Structure and Function of Plant Photoreceptors2010 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.

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.

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.

Dark Light

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

YI Wu et al. Nature 000, 1-5 (2009) doi:10.1038/nature08241

Crystallization and structural modelling of PA-Rac1.

Lov2

Jα

Rac1

Photoactive(PA)-Cdc42 binds PAK

“Lit” stateNo inhibition

Trp mutation decreases PAK binding

Adding the PAK CRIB domain inhibits binding

Photogenetics can be used to light or dark activate.

N2 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.

Structure and Function of Plant Photoreceptors2010 Annual Review by Möglich, Yang, Ayers, Moffat

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

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

YI Wu et al. Nature 000, 1-5 (2009) doi:10.1038/nature08241

Localized activation or inactivation of PA-Rac1 induces myosin-dependent migration.

YI Wu et al. Nature 000, 1-5 (2009) doi:10.1038/nature08241

Inhibition of RhoA by PA-Rac1.