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Journal of Virology, June 1999, p. 5079-5088, Vol. 73, No. 6
Unité d'Oncologie
Virale1 and Unité Rétrovirus
et Transfert Génétique,
Received 25 November 1998/Accepted 12 March 1999
Previous biochemical and genetic evidence indicated that the
functional form of retroviral integrase protein (IN) is a multimer. A
direct demonstration of IN oligomerization during the infectious cycle
was, however, missing, due to the absence of a sensitive detection method. We describe here the generation of infectious human
immunodeficiency virus type 1 (HIV-1) viral clones carrying IN protein
tagged with highly antigenic epitopes. In this setting, we could
readily visualize IN both in producer cells and in viral particles.
More interestingly, we detected IN oligomers, the formation of which
was dependent on disulfide bridges and took place inside virions.
Additionally, expression of a tagged HIV-1 IN in the absence of other
viral components resulted in almost exclusive nuclear
accumulation of the protein. Mutation of a conserved cysteine in the
proposed dimer interface determined the loss of viral infectivity, associated with a reduction of IN oligomer formation and the
redistribution of the mutated protein in the nucleus and cytoplasm.
Epitope tagging of HIV-1 IN expressed alone or in the context of a
replication-competent viral clone provides powerful tools to validate
debated issues on the implication of this enzyme in different steps of
the viral cycle.
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Oligomerization within Virions and Subcellular
Localization of Human Immunodeficiency Virus Type 1 Integrase
*
Corresponding author. Mailing address: Laboratoire de
Recherche Antivirale, IMEA/INSERM Hôpital Bichat-Claude
Bernard, 46 rue H. Huchard, 75018 Paris, France. Phone:
33-1-4025 6359. Fax: 33-1-4025 6351. E-mail:
mammano{at}bichat.inserm.fr.
Journal of Virology, June 1999, p. 5079-5088, Vol. 73, No. 6
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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