Student: Cassandra Hardy, Undergraduate Student in Chemistry, Drake University
Research Mentor: Marc Busch
Production of Recombinant UL89 from Insect Cells
Human cytomegalovirus (HCMV) is a virus in the herpes family that affects a large percentage of the U.S. population. Stress, for example spaceflight, can weaken the immune system resulting in the reactivation of latent HCMV. Currently, all of the pharmacological treatments available work by inhibiting the viral DNA polymerase, necessitating identification of a new drug and different viral targets. Resistance to this drug has been mapped to the UL89 gene. New potential therapy (2’- deoxyribosylindole (indole) nucleoside analogs) has been identified. However, exactly how indoles work to inhibit viral replication remains unresolved. UL89 is the viral nuclease responsible for the cleavage of progeny viral genomes for packaging into new virions. In order to examine if these drugs work by inhibiting this enzymatic activity, it is necessary to generate purified recombinant pUL89.
My project focuses on production of recombinant UL89 from insect cells. Insect cells are being used because of the potential to make large quantities of protein that have similar folding and post translation modification as those seen in human cells. Currently, I am working on generating a recombinant baculovirus containing the HCMV UL89 gene that will be used for generating the recombinant protein. Once the recombinant protein is generated and purified, it will be used in in vitro assays in the presence and absence of indoles to determine if these drugs block enzymatic activity. By identifying the mechanism of action, these new drugs can move forward and potentially provide a new treatment for reactivated CMV that occurs during space flight.