HIV-1 integrase interaction with U3 and U5 terminal sequences in vitro defined using substrates with random sequences.

The Journal of Biological Chemistry
Elena Brin, J Leis

Abstract

Successful integration of viral genome into a host chromosome depends on interaction between viral integrase and its recognition sequences. We have used a reconstituted concerted human immunodeficiency virus, type 1 (HIV-1), integration system to analyze the role of integrase (IN) recognition sequences in formation of the IN-viral DNA complex capable of concerted integration. HIV-1 integrase was presented with substrates that contained all 4 bases at 8 mismatched positions that define the inverted repeat relationship between U3 and U5 long terminal repeats (LTR) termini and at positions 17-19, which are conserved in the termini. Evidence presented indicates that positions 17-20 of the IN recognition sequences are needed for a concerted DNA integration mechanism. All 4 bases were found at each randomized position in sequenced concerted DNA integrants, although in some instances there were preferences for specific bases. These results indicate that integrase tolerates a significant amount of plasticity as to what constitutes an IN recognition sequence. By having several positions randomized, the concerted integrants were examined for statistically significant relationships between selections of bases at different positions. The r...Continue Reading

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Citations

Feb 1, 2011·Current Opinion in Structural Biology·Peter CherepanovStephen Hare
Dec 4, 2008·Methods : a Companion to Methods in Enzymology·Duane P GrandgenettSapna Sinha
Nov 19, 2008·Journal of Molecular Biology·James DolanJonathan Leis
Sep 18, 2012·Current Opinion in Immunology·Richard GabrielChristof von Kalle
Dec 1, 2009·Viruses·Jacques J KesslMamuka Kvaratskhelia
Mar 3, 2004·The Journal of Biological Chemistry·Ajaykumar VoraDuane Grandgenett

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