Enhancing protein stability with retained biological function in transgenic plants.
The final expression level of a transgene-derived protein in transgenic plants depends on transcriptional and post-transcriptional processes. Here, we focus on methods to improve protein stability without comprising biological function. We found that the four isoforms of the Arabidopsis RAD23 protein family are relatively stable. The UBA2 domain derived from RAD23a can be used as a portable stabilizing signal to prolong the half-life of two unstable transcription factors (TFs), HFR1 and PIF3. The increased stability of the TF-UBA2 fusion proteins results in an enhanced phenotype in transgenic plants compared to expression of the TF alone. Similar results were obtained for the RAD23a UBA1 domain. In addition to UBA1/2 of RAD23a, the UBA domain from the Arabidopsis DDI1 protein also increased the half-life of the unstable protein JAZ10.1, which is involved in jasmonate signaling. Taken together, our results suggest that UBA fusions can be used to increase the stability of unstable proteins for basic plant biology research as well as crop improvement.
Laboratory of Plant Molecular Biology, Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
In-Cheol
IC
Jang
Qi-Wen
QW
Niu
Shulin
S
Deng
Pingzhi
P
Zhao
Nam-Hai
NH
Chua
D000085:Acetates / Q000494:pharmacology; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D003517:Cyclopentanes / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D006207:Half-Life; D007976:Leupeptins / Q000494:pharmacology; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D054883:Oxylipins / Q000494:pharmacology; D010641:Phenotype; D010937:Plant Growth Regulators / Q000494:pharmacology; D030821:Plants, Genetically Modified; D020033:Protein Isoforms; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D036226:Seedlings; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D000085:Acetates; D029681:Arabidopsis Proteins; D051792:Basic Helix-Loop-Helix Proteins; D003513:Cycloheximide; D003517:Cyclopentanes; D004268:DNA-Binding Proteins; D007976:Leupeptins; D009687:Nuclear Proteins; D054883:Oxylipins; D010937:Plant Growth Regulators; D020033:Protein Isoforms; D011500:Protein Synthesis Inhibitors; D018160:Receptors, Cytoplasmic and Nuclear; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; C583231:DDI1 protein, Arabidopsis; C414438:At1g02340 protein, Arabidopsis; C525049:JAS1 protein, Arabidopsis; C116162:PIF3 protein, Arabidopsis; C583232:RAD23a protein, Arabidopsis; C072239:methyl jasmonate; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde