Graduate STEM Fellow Profile
IMPACT LA: Improving Minority Partnerships and Access through CISE-related Teaching
Thesis: Effects of Amino Acid Substitutions in the F Protein of Sendai virus on F Protein Cleavability and Plaque Formation of the Virus
College/University: California State University, Los Angeles
Research Advisor: Dr. Nancy McQueen
Degree Sought: Masters in Biology
Department: Department of Biology
Research Focus: Virology
Teaching Partner(s): Christine Rosser
Description of Research
Sendai virus (SeV), a member of the Paramyxoviridae family, contains a linear antisense RNA genome. Wild-type (wt) SeV causes a localized respiratory tract infection in mice, while it’s variant, F1-R causes a systemic (pantropic) infection. The pantropic phenotype of F1-R has been correlated to enhanced cleavability in the F gene mutations and to bipolar budding in the M gene mutations. Several SeV variants containing different combinations of the F and M gene mutations found in F1-R were generated through reverse genetics to examine the gene mutations required for a systemic infection. Studies with these variants suggested that one or more of the other amino acid substitutions in the fusion protein are required for the enhanced cleavability of F1-R F and the pantropic phenotype of F1-R. In this project we will test the hypothesis that amino acid substitutions in F1-R F encoding F115 and F116 (mutations adjacent to and at the cleavage site) will work in concert to allow for enhanced cleavability of F1-R F. Site-directed mutagenesis performed on pRGV-1 to insert the F1-R mutation encoding the F116 amino acid substitution (Arg to Lys) will generate pRGV-8. Reverse genetics will be used to construct the recombinant Sendai virus RGV-8 encoding both the F1-R F115 and F116 amino acid substitutions and lacking the M protein mutations. RGV-8 will then be studied in tissue culture with and without the addition of trypsin. We predict that RGV-8 will have an F protein with enhanced cleavability that will allow the virus to undergo multiple cycles of replication and form plaques without the addition of trypsin in tissue culture cells. As it lacks the F1-R M gene mutations, we do not believe that this virus will cause a systemic infection in mice. The data obtained herein will aid in identifying the required mutations in SeV that allow for the enhanced cleavability and pantropism of the variant F1-R. It will also provide further knowledge on the fusion process of other RNA viruses.
Example of how my research is integrated into my GK-12 experience
In every lesson I participate in I include briefly or extensively how it ties into the work that I do in order to help students make the connection back to my research/field or how to connect my science work with the lesson. For example, during cell lesson explained viruses are only non-cellular organisms known; during DNA explained that viruses have genomes; and during evolution explained how viruses change (evolve) to become more infective/pathogenic.