Avian sarcoma and leukemia virus (ASLV) integration in vitro: mutation or deletion of integrase (IN) recognition sequences does not prevent but only reduces the efficiency and accuracy of DNA integration.
Integrase (IN) is the enzyme responsible for provirus integration of retroviruses into the host cell genome. We used an Avian Sarcoma and Leukemia Viruses (ASLV) integration assay to investigate the way in which IN integrates substrates mutated or devoid of one or both IN recognition sequences. We found that replacing U5 by non-viral sequences (U5del) or U3 by a mutated sequence (pseudoU3) resulted in two and three fold reduction of two-ended integration (integration of the two ends from a donor DNA) respectively, but had a slight effect on concerted integration (integration of both ends at the same site of target DNA). Further, IN was still able to integrate the viral ends of the double mutant (pseudoU3/U5del) in a two-ended and concerted integration reaction. However, efficiency and accuracy (i.e. fidelity of size duplication and of end cleavage) of integration were reduced.
Moreau
Karen
K
Institut National de la Recherche Agronomique, UMR754, Lyon, F-69007, France.
Charmetant
Julie
J
Gallay
Kathy
K
Faure
Claudine
C
Verdier
Gérard
G
Ronfort
Corinne
C
D001355:Alpharetrovirus / Q000235:genetics* / Q000502:physiology*; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D005814:Genes, Viral; D066298:In Vitro Techniques; D019426:Integrases / Q000235:genetics* / Q000502:physiology*; D008954:Models, Biological; D009154:Mutation; D017384:Sequence Deletion; D016662:Virus Integration / Q000235:genetics* / Q000502:physiology*
D017931:DNA Primers; D004279:DNA, Viral; D019426:Integrases