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Originally published In Press as doi:10.1074/jbc.M201785200 on March 23, 2002

J. Biol. Chem., Vol. 277, Issue 23, 21071-21079, June 7, 2002
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Endonuclease G, a Candidate Human Enzyme for the Initiation of Genomic Inversion in Herpes Simplex Type 1 Virus*

Ke-Jung HuangDagger , Boris V. Zemelman§, and I. Robert LehmanDagger

From the Dagger  Department of Biochemistry, Beckman Center, Stanford University, Stanford California 94305

The herpes simplex virus type 1 (HSV-1) a sequence is present as a direct repeat at the two termini of the 152-kilobase viral genome and as an inverted repeat at the junction of the two unique components L and S. During replication, the HSV-1 genome undergoes inversion of L and S, producing an equimolar mixture of the four possible isomers. Isomerization is believed to result from recombination triggered by breakage at the a sequence, a recombinational hot spot. We have identified an enzyme in HeLa cell extracts that preferentially cleaves the a sequence and have purified it to near homogeneity. Microsequencing showed it to be human endonuclease G, an enzyme with a strong preference for G+C-rich sequences. Endonuclease G appears to be the only cellular enzyme that can specifically cleave the a sequence. Endonuclease G also showed the predicted recombination properties in an in vitro recombination assay. Based on these findings, we propose that endonuclease G initiates the a sequence-mediated inversion of the L and S components during HSV-1 DNA replication.

* This work was supported by National Institutes of Health Grant AI20538.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Present address: Memorial Sloan-Kettering Cancer Center, Box 205, 1275 York Ave., New York, NY 10021

To whom correspondence should be addressed: Dept. of Biochemistry, Stanford University, CA 94305. Tel.: 650-723-6164; Fax: 650-498-6254; E-mail: blehman@cmgm.stanford.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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