NSF Award
2350392
This project will shed light on how a DNA-directed RNA polymerase, normally involved in transcription of cellular DNA, is also able to recognize RNA templates. To understand how transcription polymerases can recognize RNA templates, this project aims to determine the molecular basis of a regulatory element within a well-known RNA template (potato spindle tuber viroid) that is critical for recruiting the transcription enzymes. To achieve this goal, this project will employ a combination of molecular, biochemical, and bioinformatic approaches. The knowledge gained from this project may facilitate the development of RNA-based technologies for basic research as well as applications in biomedicine and agriculture, thus benefiting society in the long term. This project will also serve as a vehicle for training undergraduate and graduate students. Particularly, undergraduate students from Delta State University, located in one of the least affluent regions in the US and lacking the necessary facility for research training, will participate in summer research annually to gain research experience and to be prepared for STEM-related careers.<br/><br/>Supported by published and preliminary data, a testable hypothesis is that potato spindle tuber viroid recruits RNA-specific transcription factor(s), thereby enhancing the efficiency of RNA polymerase II (Pol II)-catalyzed RNA-templated transcription. The RNA promoter region is defined by the binding region of a recently identified RNA-specific transcription factor, TFIIIA-7ZF. This project will map and characterize the molecular basis/structure of RNA promoters within both sense and antisense genomes of potato spindle tuber viroid and its relatives, using a combination of selective 2’-hydroxyl acylation analyzed by primer extension (SHAPE) analysis, functional mutagenesis, Pol II-based in vitro transcription assay, RNA footprinting assays, etc. The outcomes from this project will help better understand the nature of RNA promoters and potentially exploit RNA promoters for usage in basic science and its applications in agriculture. While working on this project, undergraduate students from two universities in the state of Mississippi will receive experience in basic research in molecular biology.<br/><br/>This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
