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Molecular and Cellular Biology, November 1998, p. 6265-6272, Vol. 18, No. 11
Department of Internal Medicine II/Molecular
Biology, University Hospital Freiburg, D-79106 Freiburg, Germany
Received 16 March 1998/Returned for modification 28 April
1998/Accepted 27 July 1998
The DNA genome of a hepatitis B virus is generated by reverse
transcription of the RNA pregenome. Replication initiation does not
involve a nucleic acid primer; instead, the hepadnavirus P protein
binds to the structured RNA encapsidation signal
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Formation of a Functional Hepatitis B Virus
Replication Initiation Complex Involves a Major Structural Alteration
in the RNA Template
, from which it
copies a short DNA primer that becomes covalently linked to the enzyme.
Using in vitro-translated duck hepatitis B virus (DHBV) P protein, we
probed the secondary structure of the protein-bound DHBV
RNA (D
)
and observed a marked conformational change compared to free D
RNA.
Several initiation-competent mutant RNAs with a different free-state
structure were similarly altered, whereas a binding-competent but
initiation-deficient variant was not, indicating the importance of the
rearrangement for replication initiation and suggesting a mechanistic
coupling to encapsidation.
*
Corresponding author. Mailing address: Department of
Internal Medicine II/Molecular Biology, University Hospital Freiburg, Hugstetter Str. 55, D-79106 Freiburg, Germany. Phone and Fax: 49-761-270 3507. E-mail: nassal2{at}ukl.uni-freiburg.de.
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