Role Of Interacting B-ZIP Transcription Factors In Arabidopsis Thaliana Seed Development: Use Of A Designed Dominant Negative Protein | 40676
Journal of Biotechnology & Biomaterials
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The B-ZIP motif is a long bipartite α-helix. The C-terminal half is responsible for dimerization whereas N-terminal binds
to sequence-specific DNA. The N-terminal basic region is unstructured in the absence of DNA but forms an alpha-helix
when bound to DNA. We have replaced the basic N-terminal domain with a rationally designed acidic extension. Dimerization
specificity of BZIP10, BZIP25 and BZIP53 with each other and with our designed dominant negative A-ZIP53 and its mutants
was evaluated using biochemical and biophysical assays. The acidic extension act as a DNA template and extends the alphahelical
coiled coil to N-terminal. This acidic extension heterodimerize with wild type basic region and inhibit the DNA binding
activities of wild type proteins. Circular dichroism results indicate that heterodimer of designed acidic leucine zipper and
basic leucine zipper is more stable than homodimer of wild type B-ZIPs. Gel shift experiments showed the inhibition of wild
type BZIP53, BZIP25 and BZIP10 by A-ZIP53 in equimolar concentrations. Transient transfections studies using Arabidopsis
protoplast proved the in vivo efficacies of these dominant negative proteins. Unlike siRNA and other knockout technology our
designed dominant negative heterodimerizes with all the memebers of same B-ZIP families thus overcoming the problem of
Prateek Jain has completed his Masters of Science in Plant Biotechnology from Banaras Hindu University in year 2012. He is currently working as PhD student since 2012 under the guidance of Dr. Vikas Rishi. He has qualified GATE Exam in Life Science (2011), DBT-JRF (2012), CSIR-NET (2012), ARS-NET (2012) and CSIR-UGC-JRF (2013). He has already presented s in two international conferences (7th International conference on Plant Biotechnology, Molecular Medicine & Human Health and Indraprastha International Conference on Biotechnology-2013).