a template-dependent dislocation mechanism potentiates k65r reverse transcriptase mutation development in subtype c variants of hiv-1template-dependent位错机制强化k65r c亚型变异的hiv - 1逆转录酶突变发展.pdfVIP

A Template-Dependent Dislocation Mechanism Potentiates K65R Reverse Transcriptase Mutation Development in Subtype C Variants of HIV-1 1,2,3 ´ 1,3 1 1 Dimitrios Coutsinos , Cedric F. Invernizzi , Daniela Moisi , Maureen Oliveira , Jorge L. Martinez- Cajas1,4, Bluma G. Brenner1,2,3, Mark A. Wainberg1,2,3* ´ ´ 1 McGill University AIDS Center, Lady Davis Institute for Medical Research, Sir Mortimer B. Davis Jewish General Hospital, Montreal, Quebec, Canada, 2 Departments of ´ ´ ´ ´ Microbiology and Immunology, McGill University, Montreal, Quebec, Canada, 3 Department of Medicine, McGill University, Montreal, Quebec, Canada, 4 Department of Medicine, Infectious Diseases, Queen’s University, Kingston, Ontario, Canada Abstract Numerous studies have suggested that the K65R reverse transcriptase (RT) mutation develops more readily in subtype C than subtype B HIV-1. We recently showed that this discrepancy lies partly in the subtype C template coding sequence that predisposes RT to pause at the site of K65R mutagenesis. However, the mechanism underlying this observation and the elevated rates of K65R development remained unknown. Here, we report that DNA synthesis performed with subtype C templates consistently produced more K65R-containing transcripts than subtype B templates, regardless of the subtype- origin of the RT enzymes employed. These findings confirm that the mechanism involved is template-specific and RT- independent. In addition, a pattern of DNA synthesis characteristic of site