Effect of metal ion binding on the structural stability of the hepatitis C virus RNA polymerase.

The Journal of Biological Chemistry
Ines BenzaghouMartin Bisaillon

Abstract

The RNA polymerase activity of the hepatitis C virus, a major human pathogen, has previously been shown to be supported by metal ions. In the present study, we report a systematic analysis of the effect of metal ion binding on the structural stability of the hepatitis C virus RNA polymerase. Chemical and thermal denaturation assays revealed that the stability of the protein is increased significantly in the presence of metal ions. Structural analyses clearly established that metal ion binding increases hydrophobic exposure on the RNA polymerase surface. Furthermore, our denaturation studies, coupled with polymerization assays, demonstrate that the active site region of the polymerase is more sensitive to chemical denaturant than other structural scaffolds. We also report the first detailed study of the thermodynamic parameters involved in the interaction between the hepatitis C virus RNA polymerase and metal ions. Finally, a mutational analysis was also performed to investigate the importance of Asp(220), Asp(318), and Asp(319) for metal ion binding. This mutational study underscores a strict requirement for each of the residues for metal binding, indicating that the active center of the HCV RNA polymerase is intolerant to virt...Continue Reading

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Citations

Feb 16, 2007·Journal of Virology·Priscilla M Van WynsberghePaul Ahlquist
Oct 22, 2008·Biochimica Et Biophysica Acta·Isabelle Bougie, Martin Bisaillon
Aug 6, 2020·Polymers·Natanel JarachSamuel Kenig
Jan 8, 2005·The Journal of Biological Chemistry·Carine FillebeenKostas Pantopoulos
Mar 10, 2021·Protein Science : a Publication of the Protein Society·Israel SilmanLev Weiner

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