RNA-binding protein-mediated translational repression of transgene expression in plants.
We have demonstrated that RNA-binding proteins from coliphages and yeast can function as translational repressors in plants. RNA sequences called translational operators were inserted at a cap-proximal position in the 5'-UTR of mRNAs of two reporter genes, gus or aroA:CP4. Translation of the reporter mRNAs was efficiently repressed when the RNA binding protein that specifically binds to its cognate operator was co-expressed. The efficiency of translational repression by RNA-binding protein positively correlated with the amount of binding protein in transformed plant cells. Detailed studies on coliphage MS2 coat protein-mediated translational repression also suggested that the efficiency of translational repression was position-dependent. A translational operator situated at the cap-proximal position was more efficient in conferring repression than one that was placed cap-distal. Translational repression can be an efficient means for regulation of transgene expression, thereby broadening current approaches for transgene regulation in plants.
Cerny
R Eric
RE
Monsanto Company, Mail Stop u4c, 800 N Lindbergh Boulevard, St. Louis, MO 63167, USA.
Qi
Youlin
Y
Aydt
Carrie M
CM
Huang
Shihshieh
S
Listello
Jennifer J
JJ
Fabbri
Brandon J
BJ
Conner
Timothy W
TW
Crossland
Lyle
L
Huang
Jintai
J
D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D019883:Alkyl and Aryl Transferases / Q000235:genetics / Q000378:metabolism; D017360:Arabidopsis / Q000235:genetics; D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D019281:Dimerization; D005786:Gene Expression Regulation; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D009875:Operator Regions, Genetic / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics*; D014176:Protein Biosynthesis / Q000235:genetics*; D011523:Protoplasts / Q000166:cytology / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019076:Transgenes / Q000235:genetics*; D003313:Zea mays / Q000166:cytology / Q000235:genetics
D036022:Capsid Proteins; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D019883:Alkyl and Aryl Transferases; D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D005966:Glucuronidase