TY - JOUR
T1 - Methylation regulates hepatitis b viral protein expression
AU - Vivekanandan, Perumal
AU - Thomas, David
AU - Torbenson, Michael
N1 - Funding Information:
Received 29 February 2008; accepted 19 June 2008; electronically published 16 March 2009. Potential conflicts of interest: none reported. Financial support: National Institutes of Health (grants K08DA016370 and R01DK078686 to M.T. and grant R01DA016078 to D.T.). Reprints or correspondence: Michael Torbenson, MD, Rm. B314, 1503 E. Jefferson, the Johns Hopkins University School of Medicine, Baltimore, MD 21231 (mtorben@jhmi.edu).
PY - 2009/5/1
Y1 - 2009/5/1
N2 - Background. Hepatitis B virus (HBV)DNAhas been shown to containCpGislands that are methylated in human tissue, which suggests a role for methylation in regulating viral protein production. However, data are lacking about whether methylation regulates viral gene expression. Methods. To investigate the hypothesis that methylation of viral DNA regulates viral gene expression, unmethylated, partially methylated, and fully methylated viralDNAwas transfected into HepG2 cells. In addition, a new assay was designed that specifically identifies methylated covalently closed circular DNA (cccDNA) in human liver tissue. Results. Transfection of methylated HBV DNA led to reduced HBV mRNA levels in HepG2 cells, decreased surface and core protein expression in these cells, and decreased secretion of HBV viral proteins into the cell supernatant. These data provide direct evidence that CpG islands regulate gene transcription of HBV. Furthermore, methylated cccDNA was found in tumor and nonneoplastic human liver tissues. Finally, an in vitro equivalent of cccDNA showed decreased viral protein production in HepG2 cells after DNA methylation. Conclusion. Taken together, these data demonstrate that methylation of viral CpG islands can regulate viral protein production.
AB - Background. Hepatitis B virus (HBV)DNAhas been shown to containCpGislands that are methylated in human tissue, which suggests a role for methylation in regulating viral protein production. However, data are lacking about whether methylation regulates viral gene expression. Methods. To investigate the hypothesis that methylation of viral DNA regulates viral gene expression, unmethylated, partially methylated, and fully methylated viralDNAwas transfected into HepG2 cells. In addition, a new assay was designed that specifically identifies methylated covalently closed circular DNA (cccDNA) in human liver tissue. Results. Transfection of methylated HBV DNA led to reduced HBV mRNA levels in HepG2 cells, decreased surface and core protein expression in these cells, and decreased secretion of HBV viral proteins into the cell supernatant. These data provide direct evidence that CpG islands regulate gene transcription of HBV. Furthermore, methylated cccDNA was found in tumor and nonneoplastic human liver tissues. Finally, an in vitro equivalent of cccDNA showed decreased viral protein production in HepG2 cells after DNA methylation. Conclusion. Taken together, these data demonstrate that methylation of viral CpG islands can regulate viral protein production.
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U2 - 10.1086/597614
DO - 10.1086/597614
M3 - Article
C2 - 19301974
AN - SCOPUS:65649090562
SN - 0022-1899
VL - 199
SP - 1286
EP - 1291
JO - Journal of Infectious Diseases
JF - Journal of Infectious Diseases
IS - 9
ER -