Document Detail


Controlling the DNA cleavage activity of light-inducible chimeric endonucleases by bidirectional photo-activation.
MedLine Citation:
PMID:  22559722     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
A functional coupling of photo-sensory domains derived from photoreceptors to effector proteins is a promising strategy for engineering novel photo-responsive proteins in optogenetics. Here, we have fused the light-sensitive LOV2 domain from <italic>Avena sativa</italic> phototropin1 to the restriction enzyme PvuII to generate a genetically encoded, light-controllable endonuclease. By analyzing several LOV-PvuII fusion enzymes, variants were obtained that show a threefold difference in DNA cleavage activity, when illuminated with blue light or kept in the dark. The effect is fully reversible over multiple photo-cycles. Depending on the particular fusion interface, the LOV-PvuII variants obtained had a bidirectional polarity in photo-activation, i.e. an increased DNA cleavage activity was observed either in the dark state, with a compact folded LOV domain, or in the blue light photo-excitation state, when the LOV domain is partially unfolded.
Authors:
Benno Schierling; Alfred Pingoud
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-5-6
Journal Detail:
Title:  Bioconjugate chemistry     Volume:  -     ISSN:  1520-4812     ISO Abbreviation:  -     Publication Date:  2012 May 
Date Detail:
Created Date:  2012-5-7     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9010319     Medline TA:  Bioconjug Chem     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
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