Alternative divalent cations (Zn2+, Co2+, and Mn2+) are not mutagenic at conditions optimal for HIV-1 reverse transcriptase activity
| dc.contributor.author | Achuthan, Vasudevan | |
| dc.contributor.author | DeStefano, Jeffrey J | |
| dc.date.accessioned | 2021-08-24T15:12:03Z | |
| dc.date.available | 2021-08-24T15:12:03Z | |
| dc.date.issued | 2015-05-03 | |
| dc.description.abstract | Fidelity of DNA polymerases can be influenced by cation co-factors. Physiologically, Mg2+ is used as a co-factor by HIV reverse transcriptase (RT) to perform catalysis; however, alternative cations including Mn2+, Co2+, and Zn2+ can also support catalysis. Although Zn2+ supports DNA synthesis, it inhibits HIV RT by significantly modifying RT catalysis. Zn2+ is currently being investigated as a component of novel treatment options against HIV and we wanted to investigate the fidelity of RT with Zn2+. We used PCR-based and plasmid-based alpha complementation assays as well as steady-state misinsertion and misincorporation assays to examine the fidelity of RT with Mn2+, Co2+, and Zn2+. The fidelity of DNA synthesis by HIV-1 RT was approximately 2.5 fold greater in Zn2+ when compared to Mg2+ at cation conditions optimized for nucleotide catalysis. Consistent with this, RT extended primers with mismatched 3′ nucleotides poorly and inserted incorrect nucleotides less efficiently using Zn2+ than Mg2+. In agreement with previous literature, we observed that Mn2+ and Co2+ dramatically decreased the fidelity of RT at highly elevated concentrations (6 mM). However, surprisingly, the fidelity of HIV RT with Mn2+ and Co2+ remained similar to Mg2+ at lower concentrations that are optimal for catalysis. This study shows that Zn2+, at optimal extension conditions, increases the fidelity of HIV-1 RT and challenges the notion that alternative cations capable of supporting polymerase catalysis are inherently mutagenic. | en_US |
| dc.description.uri | https://doi.org/10.1186/s12858-015-0041-x | |
| dc.identifier | https://doi.org/10.13016/3xku-j025 | |
| dc.identifier.citation | Achuthan, V., DeStefano, J.J. Alternative divalent cations (Zn2+, Co2+, and Mn2+) are not mutagenic at conditions optimal for HIV-1 reverse transcriptase activity. BMC Biochem 16, 12 (2015). | en_US |
| dc.identifier.uri | http://hdl.handle.net/1903/27643 | |
| dc.language.iso | en_US | en_US |
| dc.publisher | Springer Nature | en_US |
| dc.relation.isAvailableAt | Cell Biology & Molecular Genetics | en_us |
| dc.relation.isAvailableAt | Digital Repository at the University of Maryland | en_us |
| dc.relation.isAvailableAt | College of Computer, Mathematical & Natural Sciences | en_us |
| dc.relation.isAvailableAt | University of Maryland (College Park, MD) | en_us |
| dc.subject | Calf Intestinal Alkaline Phosphatase | en_US |
| dc.subject | Alternative Cation | en_US |
| dc.subject | Fidelity Assay | en_US |
| dc.subject | Average Extension Rate | en_US |
| dc.subject | Mismatch Extension | en_US |
| dc.title | Alternative divalent cations (Zn2+, Co2+, and Mn2+) are not mutagenic at conditions optimal for HIV-1 reverse transcriptase activity | en_US |
| dc.type | Article | en_US |