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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
27
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pubmed:dateCreated |
2006-7-5
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pubmed:abstractText |
Inteins are naturally occurring protein elements that catalyze their own excision from within a larger protein together with the ligation of the flanking "extein" sequences. Previously we reported the directed evolution of an intein-based molecular switch in which intein splicing in yeast cells was made dependent on the cell-permeable small molecule 4-hydroxytamoxifen (4-HT). Here we show that these evolved inteins are effective means of rendering protein function and biological signaling pathway activation dependent on 4-HT in mammalian cells. We have characterized the generality, speed, and dose dependence of ligand-induced protein splicing in murine NIH3T3 cells and in human HEK293 cells. Evolved inteins were used to control in mammalian cells the function of Gli1 and a truncated form of Gli3, two transcriptional mediators of the Hedgehog signaling pathway. Finally, we show that a complex biological process such as osteoblast differentiation can be made dependent on 4-HT using the evolved intein system. Our findings suggest that evolved small-molecule-dependent inteins may serve as a general means of achieving gene-specific, dose-dependent, post-translational, and small-molecule-induced control over protein activity in mammalian systems.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-10077605,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-10375510,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-10536138,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-10559945,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-10693759,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-10742100,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-11044998,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-11403293,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-11944939,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-12148996,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-12215652,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-12742171,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-12940738,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-14723851,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-15247421,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-15772935,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-15817467,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-16015334,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-16311596,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-1660837,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-8378770,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-8604292,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-8628671,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-8707053,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-8855277,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-9183567,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-9452474,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16819890-9843687
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/4-hydroxytamoxifen,
http://linkedlifedata.com/resource/pubmed/chemical/GLI3 protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Gli protein,
http://linkedlifedata.com/resource/pubmed/chemical/Gli3 protein, mouse,
http://linkedlifedata.com/resource/pubmed/chemical/Green Fluorescent Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Kruppel-Like Transcription Factors,
http://linkedlifedata.com/resource/pubmed/chemical/Ligands,
http://linkedlifedata.com/resource/pubmed/chemical/Nerve Tissue Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Oncogene Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Tamoxifen,
http://linkedlifedata.com/resource/pubmed/chemical/Trans-Activators,
http://linkedlifedata.com/resource/pubmed/chemical/Transcription Factors
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pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
0002-7863
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
12
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pubmed:volume |
128
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
8939-46
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pubmed:dateRevised |
2010-12-3
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pubmed:meshHeading |
pubmed-meshheading:16819890-Animals,
pubmed-meshheading:16819890-Catalysis,
pubmed-meshheading:16819890-Cell Differentiation,
pubmed-meshheading:16819890-Cell Line,
pubmed-meshheading:16819890-Directed Molecular Evolution,
pubmed-meshheading:16819890-Dose-Response Relationship, Drug,
pubmed-meshheading:16819890-Green Fluorescent Proteins,
pubmed-meshheading:16819890-Humans,
pubmed-meshheading:16819890-Inteins,
pubmed-meshheading:16819890-Kruppel-Like Transcription Factors,
pubmed-meshheading:16819890-Ligands,
pubmed-meshheading:16819890-Mice,
pubmed-meshheading:16819890-Molecular Weight,
pubmed-meshheading:16819890-NIH 3T3 Cells,
pubmed-meshheading:16819890-Nerve Tissue Proteins,
pubmed-meshheading:16819890-Oncogene Proteins,
pubmed-meshheading:16819890-Osteoblasts,
pubmed-meshheading:16819890-Signal Transduction,
pubmed-meshheading:16819890-Tamoxifen,
pubmed-meshheading:16819890-Time Factors,
pubmed-meshheading:16819890-Trans-Activators,
pubmed-meshheading:16819890-Transcription Factors
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pubmed:year |
2006
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pubmed:articleTitle |
Control of transcription factor activity and osteoblast differentiation in mammalian cells using an evolved small-molecule-dependent intein.
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pubmed:affiliation |
Department of Chemistry and Chemical Biology, Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts 02138, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, Non-P.H.S.,
Research Support, Non-U.S. Gov't,
Research Support, N.I.H., Extramural
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