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Journal of Virology, October 2001, p. 9561-9570, Vol. 75, No. 20
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.20.9561-9570.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Assembly and Catalysis of Concerted Two-End Integration Events by Moloney Murine Leukemia Virus Integrase

Fan Yang and Monica J. Roth*

Department of Biochemistry, University of Medicine and Dentistry of New Jersey--- Robert Wood Johnson Medical School, Piscataway, New Jersey 08854

Received 26 January 2001/Accepted 12 July 2001

Retroviral integration results in the stable and coordinated insertion of the two termini of the linear viral DNA into the host genome. An in vitro concerted two-end integration reaction catalyzed by the Moloney murine leukemia virus (M-MuLV) integrase (IN) was used to investigate the binding and coordination of the two viral DNA ends. Comparison of the two-end integration and strand transfer assays indicates that zinc is required for efficient concerted integration utilizing plasmid DNA as target. Complementation assays using a pair of nonoverlapping integrase domains, consisting of the HHCC domain and the core/C-terminal region, yielded products containing the correct 4-base target site duplication. The efficiency of the coordinated two-end integration varied depending on the order of addition of the individual protein and DNA components in the complementation assay. Two-end integration was most efficient when the long terminal repeat (LTR) was premixed with either the target DNA or the HHCC domain. The preference for two-end integration through preincubation of the HHCC finger with the viral DNA supports the role of this domain in the recognition and/or positioning of the LTR.

* Corresponding author. Mailing address: Department of Biochemistry, University of Medicine and Dentistry of New Jersey---Robert Wood Johnson Medical School, 675 Hoes Ln., Piscataway, NJ 08854. Phone: (732) 235-5048. Fax: (732) 235-4783. E-mail: roth{at}waksman.rutgers.edu.

Journal of Virology, October 2001, p. 9561-9570, Vol. 75, No. 20
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.20.9561-9570.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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