Journal of Virology, December 2001, p. 11263-11274, Vol. 75, No. 23
Department of Microbiology and Immunology,
University of Michigan Medical School, Ann Arbor, Michigan
48109-0620
Received 26 February 2001/Accepted 21 August 2001
A Moloney murine leukemia virus-based single-replication-cycle
assay was developed to study the effects of limiting the extent of
template and primer strand complementarity on recombinogenic template
switching. This system mimicked forced copy choice recombination in
which nascent DNA transfers from the end of a donor template to an
acceptor position on the other copackaged RNA. When acceptor target
regions with different extents of complementarity to the transferring
DNA were tested, efficient recombination occurred with as few as 14 complementary nucleotides. The frequencies of correct targeting,
transfer-associated errors, mismatch extension, and transfer before
reaching the end of the donor template were determined. All four
molecular events occurred, with their proportions varying depending on
the nature of acceptor/transferring DNA complementarity. When
complementarity was severely limited, recombination was inefficient and
most products resulted from aberrant second-strand transfer rather than
from forced template switching between RNAs. Other classes of reverse
transcription products, including some that resulted from template
switching between virus and host sequences, were also observed
when homology between the acceptor and donor was limited.
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.23.11263-11274.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Effects of Limiting Homology at the Site of
Intermolecular Recombinogenic Template Switching during Moloney
Murine Leukemia Virus Replication
*
Corresponding author. Mailing address: 1150 W. Medical
Center Dr., Rm. 5641, Ann Arbor, MI 48109-0620. Phone: (734) 936-6466. Fax: (734) 764-3562. E-mail: ateles{at}umich.edu.
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