Presenting Author
Rice University
Due to my aspirations in becoming a molecular biologist, I worked for three years as a Research Assistant in the Dr. Hung-Yi Wu’s Laboratory at National Chung Hsing University when I was an undergraduate, where I focused on studying the function of coronaviral RNA elements by using quantitative real time PCR and viral RNA transfection in order to elucidate whether the RNA elements of coronavirus are imperative for coronaviral RNA synthesis. To further explore the role of 3’-poly(A) tail based on previous discovery, I utilized coronaviral RNA with 3’-poly(A) tail of varying lengths to determine whether the 3’-poly(A) tail length is important for coronaviral RNA synthesis. In my study, I discovered that coronaviral RNA with 3’-poly(A) tail fewer than 25 nucleotides long reduced the efficiency of viral RNA synthesis compared to coronaviral RNA with 3’-poly(A) tail 25 nucleotides in length. Thus, my research result indicated that the length of 3’-poly(A) tail could affect the efficiency of coronaviral RNA synthesis.
In previous study, the substitution of 3’-poly(A) tail with 3’-poly(U), (C) or (G) tail only slightly decreases the efficiency of (-)-strand synthesis. The findings indicate that in addition to the poly(A) tail, other factors acting in trans may also participate in (-)-strand synthesis. Thus, I was encouraged to look for other factors, such as poly(A) binding protein (PABP) and coronaviral nucleocapsid protein, that may be important in viral replication. To do so, I used the electrophoretic mobility shift assay (EMSA) to ensure that interactions between RNA and the nucleocapsid protein are more closely correlated with coronaviral replication than PABP. This research resulted in a published journal articles in Journal of Virology as a contributing author (Tsung-Lin Tsai, Ching-Houng Lin, Chao-Nan Lin, Chen-Yu Lo, and Hung-Yi Wu. 2018. Interplay between the poly(A) tail, poly(A)-binding protein and coronavirus nucleocapsid protein regulates gene expression of the coronavirus and host cell. Journal of Virology doi:10.1128/jvi.01162-18), and another ‘first author’ paper is published in FEBS Journal (Chen-Yu Lo, Tsung-Lin Tsai, Chao-Nan Lin, Ching-Houng Lin, and Hung-Yi Wu. 2019. Interaction of coronavirus nucleocapsid protein with the 5'- and 3'-ends of the coronavirus genome is involved in genome circularization and negative-strand RNA synthesis. FEBS Journal. doi: 0.1111/febs.14863).
I am interested in exploring the molecular basis of DNA replication machinery, and the discovery of the mechanism might lead to the development of anticancer medicines. Thus, I attend in Dr. Yang-Gao’s lab, because I am inquisitive about how polymerase may play a main role during the duplex unwinding. Notably, I used the T7 replisome to realize how the interactions between helicase and polymerase are established. These works in the Ph.D. program facilitate me to review different mechanisms between helicases and polymerases among bacteriophages as the review article in the viruses (Chen-Yu Lo, Yang Gao. DNA Helicase-Polymerase Coupling in Bacteriophage DNA Replication. Viruses. 2021 Aug 31;13(9):1739. doi: 10.3390/v13091739.), and the article in the IJMS (Chen-Yu Lo, Yang Gao. DNA Polymerase-Parental DNA Interaction Is Essential for Helicase-Polymerase Coupling during Bacteriophage T7 DNA Replication. Int J Mol Sci. 2022 Jan 25;23(3):1342. doi: 10.3390/ijms23031342.). These experiences help me to think about the central dogma of all livings and unveil how the interactions between nucleic acid and various proteins constitute as a replicative machinery.