Single Subunit RNA Polymerases: An Insight into Their Active Sites and Catalytic Mechanism | Chapter 01 | Advances and Trends in Biotechnology and Genetics Vol. 1
Aim: To analyze various single subunit DNA dependent RNA polymerases and identify conserved motifs, active site regions among them and propose a plausible mechanism of action for these polymerases using the T7 RNA polymerase as a model system. Study Design: Bioinformatics, Biochemical, Site-directed mutagenesis and X-ray crystallographic data were analyzed. Place and Duration of Study: Department of Molecular Microbiology, School of Biotechnology, Madurai Kamaraj University, Madurai – 625 021, India, from 2010 to 2013. Methodology: The advanced version of Clustal Omega was used for protein sequence analysis of various SSU DNA dependent RNA polymerases from viruses, mitochondria and chloroplasts. Along with the conserved motifs identified by the bioinformatics analysis and with the data obtained by X-ray crystallographic, biochemical and site-directed mutagenesis (SDM) were also used to confirm the possible amino acids involved in the active sites and cata...