Investigation of Staurosporine-mediated Changes in Ebola Virus Assembly and Egress

Location: Uganda

Alumni: Scott Olehnik

Faculty: Rob Stahelin

In the present study, we wished to determine whether treatment with staurosporines, specifically UCN-01 and 7OSS, could affect the phosphatidylserine (PS) concentration of the plasma membrane, and if this change in plasma membrane composition would have any effects on the ability of the Ebola matrix protein, VP40, to undergo oligomerization and budding at the membrane. We introduced EGFP-Lact-C2, D4H-mCherry, and EGFP-VP40 to HEK293 cells treated with various concentrations of the drugs UCN-01 and 7OSS. Cells were then imaged with confocal microscopy and the percent membrane fluorescence quantified using ImageJ software. Results showed that the staurosporines only decreased localization by 10 to 12 percentage points, respectively, for UCN-01 and 7OSS with the Lact-C2 and D4H constructs. Staurosporines did not delocalize Lact-C2 and D4H to the degree that was expected based upon previous studies by Maekawa and Fairn. This may be due to the fact that the drugs were obtained from two different suppliers. Further, we found that staurosporine-mediated delocalization of VP40 was possible, with a decrease of about 13 percentage points maximally for our study. Our study shows that staurosporine-mediated delocalization of PS is possible, though not to the degree that we expected. Additionally, we determined that staurosporines are able to delocalize VP40 from the membrane and affect the oligomerization and budding of the protein. 

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