Home LiteratureArticle Details
PMID: 21037589 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Rapid blue-light-mediated induction of protein interactions in living cells.

Nature methods ·Vol. 7 ·No. 12 ·2010-12-00 ·Pages 973-5

Kennedy MJ, Hughes RM, Peteya LA, Schwartz JW, Ehlers MD, Tucker CL

Abstract

Dimerizers allowing inducible control of protein-protein interactions are powerful tools for manipulating biological processes. Here we describe genetically encoded light-inducible protein-interaction modules based on Arabidopsis thaliana cryptochrome 2 and CIB1 that require no exogenous ligands and dimerize on blue-light exposure with subsecond time resolution and subcellular spatial resolution. We demonstrate the utility of this system by inducing protein translocation, transcription and Cre recombinase-mediated DNA recombination using light.

MeSH Terms
Animals Arabidopsis/genetics,metabolism,radiation effects Arabidopsis Proteins/genetics,metabolism,radiation effects Basic Helix-Loop-Helix Transcription Factors/metabolism,radiation effects Cryptochromes/metabolism,radiation effects Kinetics Light Mammals Recombinant Fusion Proteins/metabolism,radiation effects Saccharomyces cerevisiae/genetics Transcription, Genetic/radiation effects
Chemicals
Arabidopsis Proteins Basic Helix-Loop-Helix Transcription Factors CIB1 protein, Arabidopsis CRY2 protein, Arabidopsis Cryptochromes Recombinant Fusion Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Kennedy Matthew J
Department of Neurobiology, Duke University Medical Center, Durham North Carolina, USA.
Hughes Robert M
Peteya Leslie A
Schwartz Joel W
Ehlers Michael D
Tucker Chandra L
References (18)
18 references, click to expand
  1. Cryptochrome blue light photoreceptors are activated through interconversion of flavin redox states.
    J Biol Chem. 2007 Mar 30;282(13):9383-9391 PMID: 17237227
  2. Chemically induced and light-independent cryptochrome photoreceptor activation.
    Mol Plant. 2008 Jan;1(1):4-14 PMID: 20031911
  3. Spatiotemporal control of cell signalling using a light-switchable protein interaction.
    Nature. 2009 Oct 15;461(7266):997-1001 PMID: 19749742
  4. Controlling signal transduction with synthetic ligands.
    Science. 1993 Nov 12;262(5136):1019-24 PMID: 7694365
  5. Induction of protein-protein interactions in live cells using light.
    Nat Biotechnol. 2009 Oct;27(10):941-5 PMID: 19801976
  6. Syntaxin-4 defines a domain for activity-dependent exocytosis in dendritic spines.
    Cell. 2010 Apr 30;141(3):524-35 PMID: 20434989
  7. Regulation of Cre recombinase by ligand-induced complementation of inactive fragments.
    Nucleic Acids Res. 2003 Nov 1;31(21):e131 PMID: 14576331
  8. Biochemical and genetic analysis of the yeast proteome with a movable ORF collection.
    Genes Dev. 2005 Dec 1;19(23):2816-26 PMID: 16322557
  9. A light-switchable gene promoter system.
    Nat Biotechnol. 2002 Oct;20(10):1041-4 PMID: 12219076
  10. Genetically encoded photoswitching of actin assembly through the Cdc42-WASP-Arp2/3 complex pathway.
    Proc Natl Acad Sci U S A. 2008 Sep 2;105(35):12797-802 PMID: 18728185
  11. Photoexcited CRY2 interacts with CIB1 to regulate transcription and floral initiation in Arabidopsis.
    Science. 2008 Dec 5;322(5907):1535-9 PMID: 18988809
  12. Action spectrum for cryptochrome-dependent hypocotyl growth inhibition in Arabidopsis.
    Plant Physiol. 2002 Jun;129(2):774-85 PMID: 12068118
  13. Unnatural ligands for engineered proteins: new tools for chemical genetics.
    Annu Rev Biophys Biomol Struct. 2000;29:577-606 PMID: 10940260
  14. Activation of protein splicing with light in yeast.
    Nat Methods. 2008 Apr;5(4):303-5 PMID: 18272963
  15. A novel molecular recognition motif necessary for targeting photoactivated phytochrome signaling to specific basic helix-loop-helix transcription factors.
    Plant Cell. 2004 Nov;16(11):3033-44 PMID: 15486100
  16. N-terminal domain-mediated homodimerization is required for photoreceptor activity of Arabidopsis CRYPTOCHROME 1.
    Plant Cell. 2005 May;17(5):1569-84 PMID: 15805487
  17. Preparation of organotypic hippocampal slice cultures for long-term live imaging.
    Nat Protoc. 2006;1(3):1165-71 PMID: 17406399
  18. The signaling state of Arabidopsis cryptochrome 2 contains flavin semiquinone.
    J Biol Chem. 2007 May 18;282(20):14916-22 PMID: 17355959
Article Info
Journal
Nature methods
Abbr.
Nat Methods
ISSN
1548-7105
DOI
10.1038/nmeth.1524
Published
2010-12-00
Epub
2010-00-31
Pages
973-5
Language
English
Region
United States
NLM ID
101215604
PMCID
PMC3059133
Subset
IM
Grants
Howard Hughes Medical Institute · United States
NIMH NIH HHS · R01 MH064748 · United States
NIDDK NIH HHS · R01 DK081584 · United States
NIDDK NIH HHS · R01 DK81584 · United States
NINDS NIH HHS · R01 NS039402 · United States
NIDDK NIH HHS · R01 DK081584-02 · United States
External Links
PubMed source DOI: 10.1038/nmeth.1524 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2