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J. Virol., Nov 1995, 6886-6891, Vol 69, No. 11
DD Loeb and R Tian
The current model for replication of duck hepatitis B virus has reverse
transcription initiating and copying a UUAC motif within the encapsidation
signal, epsilon, near the 5' end of the RNA template. This results in
synthesis of four nucleotides of DNA. This short minus- strand DNA product
is then transferred to a complementary position, at DR1, near the 3' end of
the RNA template. Elongation of minus-strand DNA then ensues. We have
examined the transfer of minus-strand DNA during replication of duck
hepatitis B virus in cell culture. The initial aim of this work was to
examine the effect of mutations at DR1 on the transfer process. We found
that when mutations were introduced into the UUAC motif overlapping DR1,
the 5' end of minus-DNA no longer mapped to position 2537 but was shifted
two or four nucleotides. Mismatches were predicted to exist at the new
sites of elongation. Elongation from nucleotide 2537 could be restored in
these mutants by making compensatory changes in the UUAC motif within
epsilon. This finding led us to examine limitations in the shifting of the
site of transfer. When the UUAC motif in epsilon was changed to six
different tetranucleotide motifs surrounding position 2537, transfer of
minus- strand DNA shifted predictably, albeit inefficiently. Also, when
multiple UUAC motifs were introduced near DR1, the UUAC motif at nucleotide
2537 was used preferentially. Overall, our findings confirm the current
minus-strand DNA transfer model and demonstrate a marked preference for the
site of the transfer.
Copyright © 1995, American Society for Microbiology
Transfer of the minus strand of DNA during hepadnavirus replication is not invariable but prefers a specific location
McArdle Laboratory for Cancer Research, University of Wisconsin Medical School, Madison 53706, USA.
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