1. The free energy landscape of retroviral integration.
- Author
-
Vanderlinden W, Brouns T, Walker PU, Kolbeck PJ, Milles LF, Ott W, Nickels PC, Debyser Z, and Lipfert J
- Subjects
- Crystallography, X-Ray, DNA, Viral chemistry, DNA, Viral genetics, DNA, Viral metabolism, Host-Pathogen Interactions genetics, Humans, Integrases genetics, Integrases metabolism, Macromolecular Substances, Microscopy, Atomic Force, Models, Molecular, Nucleic Acid Conformation, Nucleoproteins chemistry, Nucleoproteins genetics, Nucleoproteins metabolism, Protein Multimerization, Proviruses enzymology, Retroviridae enzymology, Spumavirus enzymology, Integrases chemistry, Proviruses genetics, Retroviridae genetics, Spumavirus genetics, Virus Integration genetics
- Abstract
Retroviral integration, the process of covalently inserting viral DNA into the host genome, is a point of no return in the replication cycle. Yet, strand transfer is intrinsically iso-energetic and it is not clear how efficient integration can be achieved. Here we investigate the dynamics of strand transfer and demonstrate that consecutive nucleoprotein intermediates interacting with a supercoiled target are increasingly stable, resulting in a net forward rate. Multivalent target interactions at discrete auxiliary interfaces render target capture irreversible, while allowing dynamic site selection. Active site binding is transient but rapidly results in strand transfer, which in turn rearranges and stabilizes the intasome in an allosteric manner. We find the resulting strand transfer complex to be mechanically stable and extremely long-lived, suggesting that a resolving agent is required in vivo. more...
- Published
- 2019
- Full Text
- View/download PDF