TY - DATA T1 - The HIV-1 Integrase Monomer Induces a Specific Interaction with LTR DNA for Concerted Integration PY - 2016/02/22 AU - Krishan K. Pandey AU - Sibes Bera AU - Duane P. Grandgenett UR - https://acs.figshare.com/articles/journal_contribution/The_HIV_1_Integrase_Monomer_Induces_a_Specific_Interaction_with_LTR_DNA_for_Concerted_Integration/2584453 DO - 10.1021/bi201247f.s001 L4 - https://ndownloader.figshare.com/files/4230478 KW - 10 mM MgSO 4 KW - strand transfer inhibitors KW - OH KW - 1 mM EDTA KW - integration KW - U 5 LTR sequences KW - 21 bp KW - immunodeficiency virus type 1 KW - ODN KW - SC KW - Integrase Monomer Induces KW - U 5 DNA KW - Concerted IntegrationThe assembly mechanism KW - HIV 5 bp host site duplication KW - PFV N2 - The assembly mechanism for the human immunodeficiency virus type 1 (HIV) synaptic complex (SC) capable of concerted integration is unknown. Molecular and structural studies have established that the HIV SC and prototype foamy virus (PFV) intasome contain a tetramer of integrase (IN) that catalyzes concerted integration. HIV IN purified in the presence of 1 mM EDTA and 10 mM MgSO4 was predominately a monomer. IN efficiently promoted concerted integration of micromolar concentrations of 3′-OH recessed and blunt-ended U5 long terminal repeat (LTR) oligonucleotide (ODN) substrates (19–42 bp) into circular target DNA. Varying HIV IN to U5 DNA showed that an IN dimer:DNA end molar ratio of 1 was optimal for concerted integration. Integration activities decreased with an increasing length of the ODN, starting from the recessed 18/20 or 19/21 bp set to the 31/33 and 40/42 bp set. Under these conditions, the average fidelity for the HIV 5 bp host site duplication with recessed and blunt-ended substrates was 56%. Modifications of U5 LTR sequences beyond 21 bp from the terminus on longer DNA (1.6 kb) did not alter the ∼32 bp DNaseI protective footprint, suggesting viral sequences beyond 21 bp were not essential for IN binding. The results suggest IN binds differentially to an 18/20 bp than to a 40/42 bp ODN substrate for concerted integration. The HIV IN monomer may be a suitable candidate for attempting crystallization of an IN–DNA complex in the absence or presence of strand transfer inhibitors. ER -