Genome replication, virion secretion, and e antigen expression of naturally occurring hepatitis B virus core promoter mutants

Sameer Parekh, Fabien Zoulim, Sang Hoon Ahn, Adrienne Tsai, Jisu Li, Shigenobu Kawai, Nasser Khan, Christian Trépo, Jack Wands, Shuping Tong

Research output: Contribution to journalArticle

205 Citations (Scopus)

Abstract

The core promoter mutants of hepatitis B virus (HBV) emerge as the dominant viral population at the late HBeAg and the anti-HBe stages of HBV infection, with the A1762T/G1764A substitutions as the hotspot mutations. The double core promoter mutations were found by many investigators to moderately enhance viral genome replication and reduce hepatitis B e antigen (HBeAg) expression. A much higher replication capacity was reported for a naturally occurring core promoter mutant implicated in the outbreak of fulminant hepatitis, which was caused by the neighboring C1766T/T1768A mutations instead. To systemically study the biological properties of naturally occurring core promoter mutants, we amplified full-length HBV genomes by PCR from sera of HBeAg+ individuals infected with genotype A. All 12 HBV genomes derived from highly viremic sera (5 × 109 to 5.7 × 109 copies of viral genome/ml) harbored wild-type core promoter sequence, whereas 37 of 43 clones from low-viremia samples (0.2 × 107 to 4.6 × 107 copies/ml) were core promoter mutants. Of the 11 wild-type genomes and 14 core promoter mutants analyzed by transfection experiments in human hepatoma cell lines, 6 core promoter mutants but none of the wild-type genomes replicated at high levels. All had 1762/ 1764 mutations and an additional substitution at position 1753 (T to C), at position 1766 (C to T), or both. Moreover, these HBV clones varied greatly in their ability to secrete enveloped viral particles irrespective of the presence of core promoter mutations. High-replication clones with 1762/1764/1766 or 1753/1762/1764/1766 mutations expressed very low levels of HBeAg, whereas high-replication clones with 1753/1762/1764 triple mutations expressed high levels of HBeAg. Experiments with site-directed mutants revealed that both 1762/1764/1766 and 1753/1762/1764/1766 mutations conferred significantly higher viral replication and lower HBeAg expression than 1762/1764 mutations alone, whereas the 1753/1762/1764 triple mutant displayed only mild reduction in HBeAg expression similar to the 1762/1764 mutant. Thus, core promoter mutations other than those at positions 1762 and 1764 can have major impact on viral DNA replication and HBeAg expression.

Original languageEnglish
Pages (from-to)6601-6612
Number of pages12
JournalJournal of Virology
Volume77
Issue number12
DOIs
Publication statusPublished - 2003 Jun 1

Fingerprint

Hepatitis B e Antigens
Hepatitis B virus
virion
Virion
promoter regions
secretion
Genome
antigens
mutation
mutants
Mutation
genome
Clone Cells
clones
Viral Genome
hepatitis B antigens
Viremia
viremia
Viral DNA
hepatitis

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Parekh, Sameer ; Zoulim, Fabien ; Ahn, Sang Hoon ; Tsai, Adrienne ; Li, Jisu ; Kawai, Shigenobu ; Khan, Nasser ; Trépo, Christian ; Wands, Jack ; Tong, Shuping. / Genome replication, virion secretion, and e antigen expression of naturally occurring hepatitis B virus core promoter mutants. In: Journal of Virology. 2003 ; Vol. 77, No. 12. pp. 6601-6612.
@article{731fe7aec5a54a1f950323de349f2885,
title = "Genome replication, virion secretion, and e antigen expression of naturally occurring hepatitis B virus core promoter mutants",
abstract = "The core promoter mutants of hepatitis B virus (HBV) emerge as the dominant viral population at the late HBeAg and the anti-HBe stages of HBV infection, with the A1762T/G1764A substitutions as the hotspot mutations. The double core promoter mutations were found by many investigators to moderately enhance viral genome replication and reduce hepatitis B e antigen (HBeAg) expression. A much higher replication capacity was reported for a naturally occurring core promoter mutant implicated in the outbreak of fulminant hepatitis, which was caused by the neighboring C1766T/T1768A mutations instead. To systemically study the biological properties of naturally occurring core promoter mutants, we amplified full-length HBV genomes by PCR from sera of HBeAg+ individuals infected with genotype A. All 12 HBV genomes derived from highly viremic sera (5 × 109 to 5.7 × 109 copies of viral genome/ml) harbored wild-type core promoter sequence, whereas 37 of 43 clones from low-viremia samples (0.2 × 107 to 4.6 × 107 copies/ml) were core promoter mutants. Of the 11 wild-type genomes and 14 core promoter mutants analyzed by transfection experiments in human hepatoma cell lines, 6 core promoter mutants but none of the wild-type genomes replicated at high levels. All had 1762/ 1764 mutations and an additional substitution at position 1753 (T to C), at position 1766 (C to T), or both. Moreover, these HBV clones varied greatly in their ability to secrete enveloped viral particles irrespective of the presence of core promoter mutations. High-replication clones with 1762/1764/1766 or 1753/1762/1764/1766 mutations expressed very low levels of HBeAg, whereas high-replication clones with 1753/1762/1764 triple mutations expressed high levels of HBeAg. Experiments with site-directed mutants revealed that both 1762/1764/1766 and 1753/1762/1764/1766 mutations conferred significantly higher viral replication and lower HBeAg expression than 1762/1764 mutations alone, whereas the 1753/1762/1764 triple mutant displayed only mild reduction in HBeAg expression similar to the 1762/1764 mutant. Thus, core promoter mutations other than those at positions 1762 and 1764 can have major impact on viral DNA replication and HBeAg expression.",
author = "Sameer Parekh and Fabien Zoulim and Ahn, {Sang Hoon} and Adrienne Tsai and Jisu Li and Shigenobu Kawai and Nasser Khan and Christian Tr{\'e}po and Jack Wands and Shuping Tong",
year = "2003",
month = "6",
day = "1",
doi = "10.1128/JVI.77.12.6601-6612.2003",
language = "English",
volume = "77",
pages = "6601--6612",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "12",

}

Genome replication, virion secretion, and e antigen expression of naturally occurring hepatitis B virus core promoter mutants. / Parekh, Sameer; Zoulim, Fabien; Ahn, Sang Hoon; Tsai, Adrienne; Li, Jisu; Kawai, Shigenobu; Khan, Nasser; Trépo, Christian; Wands, Jack; Tong, Shuping.

In: Journal of Virology, Vol. 77, No. 12, 01.06.2003, p. 6601-6612.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Genome replication, virion secretion, and e antigen expression of naturally occurring hepatitis B virus core promoter mutants

AU - Parekh, Sameer

AU - Zoulim, Fabien

AU - Ahn, Sang Hoon

AU - Tsai, Adrienne

AU - Li, Jisu

AU - Kawai, Shigenobu

AU - Khan, Nasser

AU - Trépo, Christian

AU - Wands, Jack

AU - Tong, Shuping

PY - 2003/6/1

Y1 - 2003/6/1

N2 - The core promoter mutants of hepatitis B virus (HBV) emerge as the dominant viral population at the late HBeAg and the anti-HBe stages of HBV infection, with the A1762T/G1764A substitutions as the hotspot mutations. The double core promoter mutations were found by many investigators to moderately enhance viral genome replication and reduce hepatitis B e antigen (HBeAg) expression. A much higher replication capacity was reported for a naturally occurring core promoter mutant implicated in the outbreak of fulminant hepatitis, which was caused by the neighboring C1766T/T1768A mutations instead. To systemically study the biological properties of naturally occurring core promoter mutants, we amplified full-length HBV genomes by PCR from sera of HBeAg+ individuals infected with genotype A. All 12 HBV genomes derived from highly viremic sera (5 × 109 to 5.7 × 109 copies of viral genome/ml) harbored wild-type core promoter sequence, whereas 37 of 43 clones from low-viremia samples (0.2 × 107 to 4.6 × 107 copies/ml) were core promoter mutants. Of the 11 wild-type genomes and 14 core promoter mutants analyzed by transfection experiments in human hepatoma cell lines, 6 core promoter mutants but none of the wild-type genomes replicated at high levels. All had 1762/ 1764 mutations and an additional substitution at position 1753 (T to C), at position 1766 (C to T), or both. Moreover, these HBV clones varied greatly in their ability to secrete enveloped viral particles irrespective of the presence of core promoter mutations. High-replication clones with 1762/1764/1766 or 1753/1762/1764/1766 mutations expressed very low levels of HBeAg, whereas high-replication clones with 1753/1762/1764 triple mutations expressed high levels of HBeAg. Experiments with site-directed mutants revealed that both 1762/1764/1766 and 1753/1762/1764/1766 mutations conferred significantly higher viral replication and lower HBeAg expression than 1762/1764 mutations alone, whereas the 1753/1762/1764 triple mutant displayed only mild reduction in HBeAg expression similar to the 1762/1764 mutant. Thus, core promoter mutations other than those at positions 1762 and 1764 can have major impact on viral DNA replication and HBeAg expression.

AB - The core promoter mutants of hepatitis B virus (HBV) emerge as the dominant viral population at the late HBeAg and the anti-HBe stages of HBV infection, with the A1762T/G1764A substitutions as the hotspot mutations. The double core promoter mutations were found by many investigators to moderately enhance viral genome replication and reduce hepatitis B e antigen (HBeAg) expression. A much higher replication capacity was reported for a naturally occurring core promoter mutant implicated in the outbreak of fulminant hepatitis, which was caused by the neighboring C1766T/T1768A mutations instead. To systemically study the biological properties of naturally occurring core promoter mutants, we amplified full-length HBV genomes by PCR from sera of HBeAg+ individuals infected with genotype A. All 12 HBV genomes derived from highly viremic sera (5 × 109 to 5.7 × 109 copies of viral genome/ml) harbored wild-type core promoter sequence, whereas 37 of 43 clones from low-viremia samples (0.2 × 107 to 4.6 × 107 copies/ml) were core promoter mutants. Of the 11 wild-type genomes and 14 core promoter mutants analyzed by transfection experiments in human hepatoma cell lines, 6 core promoter mutants but none of the wild-type genomes replicated at high levels. All had 1762/ 1764 mutations and an additional substitution at position 1753 (T to C), at position 1766 (C to T), or both. Moreover, these HBV clones varied greatly in their ability to secrete enveloped viral particles irrespective of the presence of core promoter mutations. High-replication clones with 1762/1764/1766 or 1753/1762/1764/1766 mutations expressed very low levels of HBeAg, whereas high-replication clones with 1753/1762/1764 triple mutations expressed high levels of HBeAg. Experiments with site-directed mutants revealed that both 1762/1764/1766 and 1753/1762/1764/1766 mutations conferred significantly higher viral replication and lower HBeAg expression than 1762/1764 mutations alone, whereas the 1753/1762/1764 triple mutant displayed only mild reduction in HBeAg expression similar to the 1762/1764 mutant. Thus, core promoter mutations other than those at positions 1762 and 1764 can have major impact on viral DNA replication and HBeAg expression.

UR - http://www.scopus.com/inward/record.url?scp=0037942914&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037942914&partnerID=8YFLogxK

U2 - 10.1128/JVI.77.12.6601-6612.2003

DO - 10.1128/JVI.77.12.6601-6612.2003

M3 - Article

C2 - 12767980

AN - SCOPUS:0037942914

VL - 77

SP - 6601

EP - 6612

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 12

ER -