The NTPase/helicase domain of hepatitis C virusnonstructural protein 3 inhibits protein kinase C inependently of its NTPase activity

• Autorzy: Hartjen P. , Hochst B. , Heim D. , Von Der Kammer H. , Lucke J. , Reinholz M. , Baier A. , Smeets R. , Wege H. , Borowski P. , Schulze zur Wiesch J.
• Języki publikacji: EN

Abstrakty

EN

Helicase motif VI is a short arginine-rich motif within the NTPase/helicase domain of the non-structural protein 3 (NS3) of the hepatitis C virus (HCV). We previously demonstrated that it reduces the catalytic activity and intracellular shuttling of protein kinase C (PKC). Thus, NS3-mediated PKC inhibition may be involved in HCV-associated hepatocellular carcinoma (HCC). In this study, we expand on our earlier results, which were obtained in experiments with short fragments of NS3, to show for the first time that the catalytically active, longer C-terminal NTPase/helicase of NS3 acts as a potent PKC inhibitor in vitro. PKC inhibition assays with the NTPase-inactive mutant NS3h-D1316A revealed a mixed type kinetic inhibition pattern. A broad range of 11 PKC isotypes was tested and all of the PKC isotypes were inhibited with IC50-values in the low micromolar range. These findings were confirmed for the wild-type NTPase/helicase domain in a non-radiometric PKC inhibition assay with ATP regeneration to rule out any effect of ATP hydrolysis caused by its NTPase activity. PKCα was inhibited with a micromolar IC50 in this assay, which compares well with our result for NS3h-D1316A (IC50 = 0.7 μM). In summary, these results confirm that catalytically active NS3 NTPase/helicase can act in an analogous manner to shorter NS3 fragments as a pseudosubstrate inhibitor of PKC.

• Wydawca: -
• Czasopismo: Cellular and Molecular Biology Letters
• Rocznik: 2013
• Tom: 18
• Numer: 3
• Opis fizyczny: p.447-458,fig.,ref.

Twórcy

autor

Hartjen P.

• Department of Virology, Bernhard-Nocht-Institute for Tropical Medicine, Bernhard-Nocht-Strasse 74, 20359 Hamburg, Germany
• I Department of Internal Medicine, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20-251 Hamburg, Germany
• Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

autor

Hochst B.

• Department of Virology, Bernhard-Nocht-Institute for Tropical Medicine, Bernhard-Nocht-Strasse 74, 20359 Hamburg, Germany
• Institutes of Molecular Medicine and Experimental Immunology, University Hospital Bonn, Sigmund-Freud-Str. 25, 53105 Bonn, Germany

autor

Heim D.

• I Department of Internal Medicine, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20-251 Hamburg, Germany

autor

Von Der Kammer H.

• Department of Virology, Bernhard-Nocht-Institute for Tropical Medicine, Bernhard-Nocht-Strasse 74, 20359 Hamburg, Germany

autor

Lucke J.

• Department of Virology, Bernhard-Nocht-Institute for Tropical Medicine, Bernhard-Nocht-Strasse 74, 20359 Hamburg, Germany

autor

Reinholz M.

• Department of Virology, Bernhard-Nocht-Institute for Tropical Medicine, Bernhard-Nocht-Strasse 74, 20359 Hamburg, Germany

autor

Baier A.

• Department of Molecular Biology, The John Paul II Catholic University of Lublin, Konstantynow 1I, 20-708 Lublin, Poland

autor

Smeets R.

• Department of Oral and Maxillofacial Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

autor

Wege H.

• I Department of Internal Medicine, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20-251 Hamburg, Germany

autor

Borowski P.

• Department of Virology, Bernhard-Nocht-Institute for Tropical Medicine, Bernhard-Nocht-Strasse 74, 20359 Hamburg, Germany
• Department of Molecular Biology, The John Paul II Catholic University of Lublin, Konstantynow 1I, 20-708 Lublin, Poland

autor

Schulze zur Wiesch J.

• I Department of Internal Medicine, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20-251 Hamburg, Germany

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