Identification of natural and artificial DNA substrates for light-activated LOV-HTH transcription factor EL222.
Identification of natural and artificial DNA substrates for light-activated LOV-HTH transcription factor EL222.
复制标题
DOI：
10.1021/bi301306t
复制
发表时间：
2012-12-18
期刊：
影响因子：
2.9
通讯作者：
Gardner KH
中科院分区：
文献类型：
--
作者：
Rivera-Cancel G;Motta-Mena LB;Gardner KH
Light-oxygen-voltage (LOV) domains serve as the photosensory modules for a wide range of plant and bacterial proteins, conferring blue light dependent regulation to effector activities as diverse as enzymes and DNA binding. LOV domains can also be engineered into a variety of exogenous targets, enabling similar regulation for new protein-based reagents. Common to these proteins is the ability for LOV domains to reversibly form a photochemical adduct between an internal flavin chromophore and the surrounding protein, using this to trigger conformational changes that affect output activity. Using the Erythrobacter litoralis protein EL222 model system which links LOV regulation to a helix-turn-helix (HTH) DNA binding domain, we demonstrated that the LOV domain binds and inhibits the HTH domain in the dark, releasing these interactions upon illumination [Nash et al. (2011) Proc. Natl. Acad. Sci. USA 108, 9449–9454]. Here we combine genomic and in vitro selection approaches to identify optimal DNA binding sites for EL222. Within the bacterial host, we observe binding several genomic sites using a 12 bp sequence consensus that is also found by in vitro selection methods. Sequence-specific alterations in the DNA consensus reduce EL222-binding affinity in a manner consistent with the expected binding mode: a protein dimer binding to two repeats. Finally, we demonstrate the light-dependent activation of transcription of two genes adjacent to an EL222 binding site. Taken together, these results shed light on the native function of EL222 and provide useful reagents for further basic and applications research of this versatile protein.
登录
查看更多内容
影响因子：
2.9
作者：
Harper, SM;Christie, JM;Gardner, KH
通讯作者：
Gardner, KH
DOI：
10.1093/bioinformatics/btp472
发表时间：
2009-10-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Nicol JW;Helt GA;Blanchard SG Jr;Raja A;Loraine AE
通讯作者：
Loraine AE
影响因子：
16
作者：
Mouw, Kent W.;Rowland, Sally-J.;Gajjar, Mark M.;Boocock, Martin R.;Stark, W. Marshall;Rice, Phoebe A.
通讯作者：
Rice, Phoebe A.
影响因子：
4.8
作者：
BARAK, Y;COHENFIX, O;LIVNEH, Z
通讯作者：
LIVNEH, Z
影响因子：
3.2
作者：
Krauss, Ulrich;Minh, Bui Quang;Jaeger, Karl-Erich
通讯作者：
Jaeger, Karl-Erich