Journal
JOURNAL OF MOLECULAR BIOLOGY
Volume 342, Issue 1, Pages 57-71Publisher
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.jmb.2004.07.012
Keywords
ICP8; UL12; herpes; recombinase; electron microscopy
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Funding
- NCI NIH HHS [P01 CA019014, CA19014] Funding Source: Medline
- NIAID NIH HHS [R01 AI021747, R01 AI037549, R01 AI069136, R37 AI021747] Funding Source: Medline
- NIGMS NIH HHS [GM31819, R01 GM031819] Funding Source: Medline
- PHS HHS [A121747, A137549] Funding Source: Medline
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The replication of herpes simplex virus type 1 (HSV-1) is associated with a high degree of homologous recombination, which is likely to be mediated, in part, by HSV-1-encoded proteins. We have previously shown that the HSV-1 encoded ICP8 protein and alkaline nuclease UL12 are capable of catalyzing an in vitro strand-exchange reaction. Here, we show, by electron microscopy, that the products of the strand exchange reaction between linear double-stranded DNA and circular single-stranded DNA consist of the expected joint molecule forms: sigma, alpha, and gapped circles. Other exonucleases, such as lambda Red a, which, like UL12, digests 5'-3', as well as Escherichia coli exonuclease III (ExoIII), which digests 3'-5', could substitute for UL12 in the strand exchange reaction by providing a resected DNA end. ICP8 generated the same intermediates and strand exchange products when the double-stranded DNA substrate was preresected by any of the nucleases. Using substrates with large regions of non-homology we found that pairing by ICP8 could be initiated from the middle of a DNA molecule and did not require a homologous end. In this reaction, the resection of a DNA end by the nuclease is required to reveal homologous sequences capable of being paired by ICP8. This study further illustrates the complexity of the multi-functional ICP8 protein. (C) 2004 Elsevier Ltd. All rights reserved.
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