Abstract
Two critical events are the signature of the life cycle of retroviruses (1). The first is reverse transcription, whereby the single-stranded RNA genome of the retrovirus is copied into double-stranded DNA. The second of these events is integration, whereby this viral DNA is inserted into a chromosome of the host cell, establishing what is known as the proviral state. The proviral state is required for efficient replication of retroviruses. This crucial second event is catalyzed by the integrase enzyme. Retroviruses encode the integrase at the 3′ end of the pol gene. Integrase is generated by the retroviral protease as a proteolytic cleavage product of the gag-pol fusion protein precursor, and is contained in the virus particle. During viral infection, integrase catalyzes the excision of the last two nucleotides from each 3′ end of the linear viral DNA, leaving the terminal dinucleotide CA-3P-OH at these recessed 3′ ends. This activity is referred to as the 3′-processing or dinucleotide cleavage. After transport to the nucleus as a nucleoprotein complex (“preintegration complex”), integrase catalyzes a DNA strand transfer reaction (3′-end joining) involving the nucleophilic attack of these ends on a host chromosome. Completion of the integration process requires removal of the two unpaired nucleotides at the 5′ ends of the viral DNA and gap repair reactions that are thought to be accomplished by cellular enzymes. For recent reviews, see Andrake and Skalka (2) and Rice et al. (3).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Varmus, H. E. and Brown, P. O. (1989) in Mobile DNA (Berg, D. and Howe, M., eds.), Am. Soc. Microbiol., Washington, DC, pp. 53–108.
Andrake, M. D. and Skalka, A. M. (1996) J. Biol. Chem. 271, 19,633–19,636.
Rice, P., Craigie, R., and Davies, D. R. (1996) Curr. Opin. Struct. Biol. 6, 76–83.
Cushman, M., Golebiewski, W. M., Pommier, Y., Mazumder, A., Reymen, D., De Clerq, E., Graham, L., and Rice, W. G. (1995) J. Med. Chem. 38, 443–452.
LaFemina, R. L., Graham, P. L., LeGrow, K., Hastings, J. C., Wolfe, A., Young, S. D., Emini, E. A., and Hazuda, D. J. (1995) Antimicrob. Agents Chemother. 39, 320–324.
Fesen, M. R., Kohn, K. W., Leteurtre, F., and Pommier, Y. (1993) Proc. Natl. Acad. Sci. USA 90, 2399–2403.
Robinson, W. E., Jr., Reinecke, M. G., Abdel-Malek., S., Jia, Q., and Chow, S. A. (1996) Proc. Natl. Acad. Sci. USA 93, 6326–6331.
Bouziane, M., Cherny, D. I., Mouscadet, J. F., and Auclair, C. (1996) J. Biol. Chem. 271, 10,359–10,364.
Mazumder, A., Wang, S., Neamati, N., Nicklaus, M., Sunder, S., Chen, J., Milne, G. W. A., Rice, W. G., Burke, T. R. J., and Pommier, Y. (1996) J. Med. Chem. 39, 2472–2481.
Mazumder, A., Neamati, N., Ojwang, J. O., Sunder, S., Rando, R. F., and Pommier, Y. (1996) Biochemistry 35, 13,762–13,771.
Farnet, C. M., Wang, B., Lipford, J. R., and Bushmarn, F. D. (1996) Proc. Natl. Acad. Sci. USA 93, 9742–9747.
Mazumder, A., Engelman, A., Craigie, R., Fesen, M., and Pommier, Y. (1994) Nucleic Acids Res. 22, 1037–1043.
Mazumder, A., Gupta, M., and Pommier, Y. (1994) Nucleic Acids Res. 22, 4441–4448.
Mazumder, A., and Pommier, Y. (1995) Nucleic Acids Res. 23, 2865–2871.
Mazumder, A., Neamati, N., Pilon, A., Sunder, S., and Pommier, Y. (1996) J. Biol. Chem. 271, 27,330–27,338.
Craigie, R., Fujiwara, T., and Bushman, F. (1990) Cell 62, 829–837.
Katz, R. A., Merkel, G., Kulkosky, J., Leis, J., and Skalka, A. M. (1990) Cell 63, 87–95.
Engelman, A., Mizuuchi, K., and Craigie, R. (1991) Cell 67, 1211–1221.
Vink, C., Yeheskiely, E., van der Marel, G. A., van Boom, J. H., and Plasterk, R. H. (1991) Nucleic Acids Res. 19, 6691–6698.
Chow, S. A., Vincent, K. A., Ellison, V., and Brown, P. O. (1992) Science 255, 723–726.
Sherman, P. A., Dickson, M. L., and Fyfe, J. A. (1992) J. Virol. 66, 3593–3601.
Bushman, F. D., Engelman, A., Palmer, I., Wingfield, P., and Craigie, R. (1993) Proc. Natl. Acad. Sci. USA 90, 3428–3432.
Chow, S. A. and Brown, P. O. (1994) J. Virol. 68, 7869–7878.
van Den Ent, F. M. I., Vink, C., and Plasterk, R. H. A. (1994) J. Virol. 68, 7825–7832.
Yoshinaga, T., Kimura-Ohtani, Y., and Fujiwara, T. (1994) J. Virol. 68, 5690–5697.
Engelman, A., Hickman, A. B., and Craigie, R. (1994) J. Virol. 68, 5911–5917.
Jenkins, T. M., Engelman, A., Ghirlando, R., and Craigie, R. (1996) J. Biol. Chem. 271, 7712–7718.
Engelman, A. and Craigie, R. (1995) J. Virol. 69, 5908–5911.
Fesen, M., Pommier, Y., Leteurtre, F., Hiroguchi, S., Yung, J., and Kohn, K. W. (1994) Biochem. Pharmacol. 48, 595–608.
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2000 Humana Press Inc.
About this protocol
Cite this protocol
Mazumder, A., Neamati, N., Sunder, S., Owen, J., Pommier, Y. (2000). Retroviral Integrase. In: Kinchington, D., Schinazi, R.F. (eds) Antiviral Methods and Protocols. Methods in Molecular Medicine™, vol 24. Humana Press. https://doi.org/10.1385/1-59259-245-7:327
Download citation
DOI: https://doi.org/10.1385/1-59259-245-7:327
Publisher Name: Humana Press
Print ISBN: 978-0-89603-561-4
Online ISBN: 978-1-59259-245-6
eBook Packages: Springer Protocols