Requirement of Heat Shock Protein 90 for Human Hepatitis B Virus Reverse Transcriptase Function
AUTOR(ES)
Hu, Jianming
FONTE
American Society for Microbiology
RESUMO
The initiation of reverse transcription and nucleocapsid assembly in hepatitis B virus (HBV) depends on the specific recognition of an RNA signal (the packaging signal, ɛ) on the pregenomic RNA (pgRNA) by the viral reverse transcriptase (RT). RT-ɛ interaction in the duck hepatitis B virus (DHBV) was recently shown to require the molecular chaperone complex, the heat shock protein 90 (Hsp90). However, the requirement for RT-ɛ interaction in the human HBV has remained unknown due to the inability to obtain a purified RT protein active in specific ɛ binding. We now report that Hsp90 is also required for HBV RT-ɛ interaction. Inhibition of Hsp90 led to diminished HBV pgRNA packaging into nucleocapsids in cells, which depends on RT-ɛ interaction. Furthermore, using truncated HBV RT proteins purified from bacteria and five purified Hsp90 chaperone factors, we have developed an in vitro RT-ɛ binding assay. Our results demonstrate that Hsp90, in a dynamic process that was dependent on ATP hydrolysis, facilitated RT-ɛ interaction in HBV, as in DHBV. Specific ɛ binding required sequences from both the amino-terminal terminal protein and the carboxy-terminal RT domain. Only the cognate HBV ɛ, but not the DHBV ɛ, could bind the HBV RT proteins. Furthermore, the internal bulge, but not the apical loop, of ɛ was required for RT binding. The establishment of a defined in vitro reconstitution system has now paved the way for future biochemical and structural studies to elucidate the mechanisms of RT-ɛ interaction and chaperone activation.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=525004Documentos Relacionados
- Heat Shock Protein 90-Independent Activation of Truncated Hepadnavirus Reverse Transcriptase
- Hsp90 is required for the activity of a hepatitis B virus reverse transcriptase.
- Interaction between Hepatitis B Virus Core Protein and Reverse Transcriptase
- In Vitro Reconstitution of a Functional Duck Hepatitis B Virus Reverse Transcriptase: Posttranslational Activation by Hsp90
- Translation of Duck Hepatitis B Virus Reverse Transcriptase by Ribosomal Shunting