Host site selection for concerted integration by human immunodeficiency virus type-1 virions in vitro

Virology
G GoodarziD P Grandgenett

Abstract

Host site selection for full-site integration by human immunodeficiency virus type-1 (HIV-1) intergrase (IN) from nonionic detergent-lysed virions was investigated. Linear retrovirus-like DNA (469 bp) possessing 3' OH recessed long terminal repeat termini was efficiently inserted by a bimolecular donor reaction into a supercoiled DNA target (2867 bp), producing the HIV-1 5-bp host site duplication. Sequence data were analyzed from 193 donor-target recombinants obtained from the linear 3.8-kb DNA product. The selection of host target sites appeared randomly distributed and was independent of lysis and assay conditions. The fidelity of the 5-bp duplications in comparison to other size duplications was highest (94%) with high-salt (300 mM NaCl) lysis of the virions and 60 mM NaCl for strand transfer using Mg2+ as the divalent cation. Base sequence analysis demonstrated some biases in the 5-bp duplications at the sites of strand transfer and at the immediate host sequences surrounding the duplications. In addition to the observed duplications, approximately 30% of the recombinants isolated from the linear 3.8-kb DNA product contained specific and repetitive small-size deletions. No deletions smaller that 17 bp were observed and the...Continue Reading

References

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Citations

Apr 21, 2005·Proceedings of the National Academy of Sciences of the United States of America·Duane P Grandgenett
Apr 2, 2005·Proceedings of the National Academy of Sciences of the United States of America·Alexander G Holman, John M Coffin
Dec 4, 2008·Methods : a Companion to Methods in Enzymology·Duane P GrandgenettSapna Sinha
Jun 25, 2009·Archives of Biochemistry and Biophysics·Jan SnáselIva Pichová
Nov 18, 2005·Advances in Genetics·Mary K Lewinski, Frederic D Bushman
Dec 1, 2009·Viruses·Jacques J KesslMamuka Kvaratskhelia
Nov 14, 2015·Journal of Biosciences·B K Sarkar, Chiranjib Chakraborty

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