Direct and specific chemical control of eukaryotic translation with a synthetic RNA-protein interaction

被引:33
作者
Goldfless, Stephen J. [1 ]
Belmont, Brian J. [1 ]
de Paz, Alexandra M. [1 ]
Liu, Jessica F. [1 ]
Niles, Jacquin C. [1 ]
机构
[1] MIT, Dept Biol Engn, Cambridge, MA 02139 USA
基金
美国国家科学基金会; 美国国家卫生研究院;
关键词
MESSENGER-RNA; CAENORHABDITIS-ELEGANS; GENE-EXPRESSION; SACCHAROMYCES-CEREVISIAE; IN-VIVO; INITIATION; REPRESSION; CELLS; MECHANISMS; STABILITY;
D O I
10.1093/nar/gks028
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Sequence-specific RNA-protein interactions, though commonly used in biological systems to regulate translation, are challenging to selectively modulate. Here, we demonstrate the use of a chemically-inducible RNA-protein interaction to regulate eukaryotic translation. By genetically encoding Tet Repressor protein (TetR)-binding RNA elements into the 5'-untranslated region (5'-UTR) of an mRNA, translation of a downstream coding sequence is directly controlled by TetR and tetracycline analogs. In endogenous and synthetic 5'-UTR contexts, this system efficiently regulates the expression of multiple target genes, and is sufficiently stringent to distinguish functional from non-functional RNA-TetR interactions. Using a reverse TetR variant, we illustrate the potential for expanding the regulatory properties of the system through protein engineering strategies.
引用
收藏
页数:12
相关论文
共 38 条
[1]   Post-transcriptional regulons coordinate the initiation and resolution of inflammation [J].
Anderson, Paul .
NATURE REVIEWS IMMUNOLOGY, 2010, 10 (01) :24-35
[2]   Rational design of a small molecule-responsive intramer controlling transgene expression in mammalian cells [J].
Auslaender, David ;
Wieland, Markus ;
Auslaender, Simon ;
Tigges, Marcel ;
Fussenegger, Martin .
NUCLEIC ACIDS RESEARCH, 2011, 39 (22) :e155
[3]   Engineering a Direct and Inducible Protein-RNA Interaction To Regulate RNA Biology [J].
Belmont, Brian J. ;
Niles, Jacquin C. .
ACS CHEMICAL BIOLOGY, 2010, 5 (09) :851-861
[4]   MESSENGER-RNAS ENCODING RIBULOSE-1,5-BISPHOSPHATE CARBOXYLASE REMAIN BOUND TO POLYSOMES BUT ARE NOT TRANSLATED IN AMARANTH SEEDLINGS TRANSFERRED TO DARKNESS [J].
BERRY, JO ;
CARR, JP ;
KLESSIG, DF .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (12) :4190-4194
[5]   Translational control of localized mRNAs: restricting protein synthesis in space and time [J].
Besse, Florence ;
Ephrussi, Anne .
NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2008, 9 (12) :971-980
[6]   Origins and Mechanisms of miRNAs and siRNAs [J].
Carthew, Richard W. ;
Sontheimer, Erik J. .
CELL, 2009, 136 (04) :642-655
[7]   REVERSAL OF CREATINE-KINASE TRANSLATIONAL REPRESSION BY 3' UNTRANSLATED SEQUENCES [J].
CHNG, JLC ;
SHOEMAKER, DL ;
SCHIMMEL, P ;
HOLMES, EW .
SCIENCE, 1990, 248 (4958) :1003-1006
[8]  
Condeelis J, 2005, BIOL CELL, V97, P97
[9]   Regulation of mRNA Translation and Stability by microRNAs [J].
Fabian, Marc Robert ;
Sonenberg, Nahum ;
Filipowicz, Witold .
ANNUAL REVIEW OF BIOCHEMISTRY, VOL 79, 2010, 79 :351-379
[10]   Molecular mechanisms of translational control [J].
Gebauer, F ;
Hentze, MW .
NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2004, 5 (10) :827-835