C-terminal substitution of HBV core proteins with those from DHBV reveals that arginine-rich 167RRRSQSPRR175 domain is critical for HBV replication.
To investigate the contributions of carboxyl-terminal nucleic acid binding domain of HBV core (C) protein for hepatitis B virus (HBV) replication, chimeric HBV C proteins were generated by substituting varying lengths of the carboxyl-terminus of duck hepatitis B virus (DHBV) C protein for the corresponding regions of HBV C protein. All chimeric C proteins formed core particles. A chimeric C protein with 221-262 amino acids of DHBV C protein, in place of 146-185 amino acids of the HBV C protein, supported HBV pregenomic RNA (pgRNA) encapsidation and DNA synthesis: 40% amino acid sequence identity or 45% homology in the nucleic-acid binding domain of HBV C protein was sufficient for pgRNA encapsidation and DNA synthesis, although we predominantly detected spliced DNA. A chimeric C protein with 221-241 and 251-262 amino acids of DHBV C, in place of HBV C 146-166 and 176-185 amino acids, respectively, could rescue full-length DNA synthesis. However, a reciprocal C chimera with 242-250 of DHBV C ((242)RAGSPLPRS(250)) introduced in place of 167-175 of HBV C ((167)RRRSQSPRR(175)) significantly decreased pgRNA encapsidation and DNA synthesis, and full-length DNA was not detected, demonstrating that the arginine-rich (167)RRRSQSPRR(175) domain may be critical for efficient viral replication. Five amino acids differing between viral species (underlined above) were tested for replication rescue; R169 and R175 were found to be important.
Department of Microbiology, Ajou University School of Medicine, Woncheon-dong, Suwon, Korea.
Jaesung
J
Jung
Hee-Young
HY
Kim
Taeyeung
T
Kim
Bo-Hye
BH
Shin
Gil-Soon
GS
Park
Sun
S
Park
Yong-Joon
YJ
Chwae
Ho-Joon
HJ
Shin
Kyongmin
K
Kim
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001120:Arginine / Q000737:chemistry*; D002460:Cell Line; D004261:DNA Replication; D004279:DNA, Viral; D015611:Hepatitis B Virus, Duck / Q000502:physiology*; D006515:Hepatitis B virus / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D050506:Mutant Chimeric Proteins / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D012313:RNA / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism; D014758:Viral Core Proteins / Q000737:chemistry*; D014779:Virus Replication
D004279:DNA, Viral; D050506:Mutant Chimeric Proteins; D012367:RNA, Viral; D014758:Viral Core Proteins; C100808:pgRNA; D012313:RNA; D001120:Arginine