Author

Chalasani

Date of Award

4-1994

Degree Name

Master of Science

Department

Biological Sciences

First Advisor

Dr. Karim Essani

Second Advisor

Dr. Robert Eisenberg

Third Advisor

Dr. Louis Toledo-Pereyera

Access Setting

Masters Thesis-Open Access

Abstract

DNA methylation has a profound effect on the expression of a number of eukaryotic genes. This study demonstrates that an Epstein-Barr virus (EBV) positive Burkitt's lymphoma cell line, Raji, contains the gene encoding gp350/220 but is not expressed. Frog virus 3 (FV3), an iridovirus, has the ability to overcome the inhibitory effect of DNA methylation. Fusion of FV3-infected cells with Raji cells resulted in expression of gp350/220, suggesting that gp350/220 is regulated by DNA methylation. Further evidence for this hypothesis was obtained by transfecting B95.8 cells with HpaII/HhaI methylated plasmid pLATE-CAT, containing the gp350/220 promoter. Southern blot analyses and a modified polymerase chain reaction revealed that the gp350/220 promoter is not methylated in Raji cells. These results, taken together, provide evidence that gp350/220 in Raji cells is regulated by a methylation-sensitive transactivator. Since gp350/220 is a neutralizing antigen, these results imply that perhaps EBV takes advantage of DNA methylation to evade the host immune system.

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