TY - JOUR
T1 - Differential assembly of Hepatitis B Virus core protein on single- and double-stranded nucleic acid suggest the dsDNA-filled core is spring-loaded
AU - Dhason, Mary S.
AU - Wang, Joseph C.Y.
AU - Hagan, Michael F.
AU - Zlotnick, Adam
N1 - Funding Information:
We wish to thank Drs Chao Chen, J. Zachary Porterfield, David Nickens, and Sarah Katen for critical discussions. Electron microscopy was performed at the IU molecular biology institute; dynamic light scattering experiments were performed at the IU Physical Biochemistry Instrumentation Facility. MSD, JC-YW, and AZ were supported by NIH R01-AI077688 to AZ; MFH was supported by NIH R01-AI080791 and Burroughs-Wellcome Fund 2012 Collaborative Research Travel Grant.
PY - 2012/8/15
Y1 - 2012/8/15
N2 - Hepatitis B Virus (HBV) cores assemble on viral RNA, which is reverse transcribed within the core to the partially dsDNA genome of mature HBV. However, constraining dsDNA, a stiff polymer, within a core necessarily requires far greater capsid stability than constraining ssRNA. We hypothesized that, unlike ssRNA, dsDNA would be a poor substrate for assembly. We examined titrations of ssDNA and dsDNA with purified HBV core protein, Cp183, by EMSA, EM, DLS, and etheno-DNA fluorescence. Cp183 bound ssDNA with high affinity to form virus-like capsids. However, Cp183 bound dsDNA poorly, forming a mixture of irregular complexes. Nonetheless, we observed some normal cores in dsDNA assembly reactions, indicating that the energy required to bend DNA could be similar to the protein-protein association energy. This similarity of energies suggests that dsDNA stresses mature HBV cores, in agreement with calculation, which may be the basis for the virus maturation signal and DNA release.
AB - Hepatitis B Virus (HBV) cores assemble on viral RNA, which is reverse transcribed within the core to the partially dsDNA genome of mature HBV. However, constraining dsDNA, a stiff polymer, within a core necessarily requires far greater capsid stability than constraining ssRNA. We hypothesized that, unlike ssRNA, dsDNA would be a poor substrate for assembly. We examined titrations of ssDNA and dsDNA with purified HBV core protein, Cp183, by EMSA, EM, DLS, and etheno-DNA fluorescence. Cp183 bound ssDNA with high affinity to form virus-like capsids. However, Cp183 bound dsDNA poorly, forming a mixture of irregular complexes. Nonetheless, we observed some normal cores in dsDNA assembly reactions, indicating that the energy required to bend DNA could be similar to the protein-protein association energy. This similarity of energies suggests that dsDNA stresses mature HBV cores, in agreement with calculation, which may be the basis for the virus maturation signal and DNA release.
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U2 - 10.1016/j.virol.2012.04.012
DO - 10.1016/j.virol.2012.04.012
M3 - Article
C2 - 22595445
AN - SCOPUS:84861836672
SN - 0042-6822
VL - 430
SP - 20
EP - 29
JO - Virology
JF - Virology
IS - 1
ER -