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Regulation of the biochemical function of motif VI of HCV NTPase/helicase by the conserved Phe-loop

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Regulation of the biochemical function of motif VI of HCV NTPase/helicase by the conserved Phe-loop. / Hartjen, Philip; Medom, Berthe Kamdem; Reinholz, Michael; Borowski, Peter; Baier, Andrea.

In: BIOCHIMIE, Vol. 91, No. 2, 02.2009, p. 252-60.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Hartjen, P, Medom, BK, Reinholz, M, Borowski, P & Baier, A 2009, 'Regulation of the biochemical function of motif VI of HCV NTPase/helicase by the conserved Phe-loop', BIOCHIMIE, vol. 91, no. 2, pp. 252-60. https://doi.org/10.1016/j.biochi.2008.09.007

APA

Hartjen, P., Medom, B. K., Reinholz, M., Borowski, P., & Baier, A. (2009). Regulation of the biochemical function of motif VI of HCV NTPase/helicase by the conserved Phe-loop. BIOCHIMIE, 91(2), 252-60. https://doi.org/10.1016/j.biochi.2008.09.007

Vancouver

Hartjen P, Medom BK, Reinholz M, Borowski P, Baier A. Regulation of the biochemical function of motif VI of HCV NTPase/helicase by the conserved Phe-loop. BIOCHIMIE. 2009 Feb;91(2):252-60. https://doi.org/10.1016/j.biochi.2008.09.007

Bibtex

@article{e9a3c7981c83449b82f656e42e316905,
title = "Regulation of the biochemical function of motif VI of HCV NTPase/helicase by the conserved Phe-loop",
abstract = "In previous works, we demonstrated a potent inhibition of diverse protein kinase C (PKC) functions by a fragment of nonstructural protein 3 (NS3) of hepatitis C virus (HCV), mainly mediated by the Arg-rich amino acid motif HCV(1487-1500). This sequence is localized on the surface of Domain 2 of the NS3 NTPase/helicase in direct vicinity to a flexible loop that is localized between Val1458 and Thr1476. Here, we assessed the regulation of the accessibility of the Arg-rich amino acid motif for PKC by this loop, using two variants of domain 2. The first construct, termed NS3d2Delta, comprises the complete domain, HCV(1361-1503), devoid the loop. The second variant, NS3d2wt corresponds to wild type domain 2. The results indicated an enhanced inhibitory potential of NS3d2Delta towards rat brain PKC and towards the majority of PKC isoforms. This effect and the accompanying change of the mode of inhibition from a mixed mode, exerted by NS3d2wt to a competitive mode, exerted by NS3d2Delta are caused by the deletion of the loop. Accordingly, the presence of the intact loop abolished the binding of domain 2 to the tailed duplex RNA used as helicase substrate, without affecting the binding of dsDNA. Furthermore, a direct competition of dsRNA and PKC for the same binding site HCV(1487-1500), could be documented. The binding of dsRNA to NS3d2Delta previously overlaid with PPKCalpha was reduced to 30% and completely abolished in case of NS3d2Delta overlaid with cAMP-dependent protein kinase A (PKA).",
keywords = "Amino Acid Motifs, Amino Acid Sequence, Animals, Binding Sites, Conserved Sequence, Escherichia coli, Hepacivirus, Models, Molecular, Molecular Sequence Data, Nucleoside-Triphosphatase, Plasmids, Protein Isoforms, Protein Kinase C, Protein Kinase C-alpha, Protein Structure, Secondary, Protein Structure, Tertiary, RNA Helicases, Rats, Recombinant Proteins, Sequence Homology, Amino Acid, Viral Nonstructural Proteins",
author = "Philip Hartjen and Medom, {Berthe Kamdem} and Michael Reinholz and Peter Borowski and Andrea Baier",
year = "2009",
month = feb,
doi = "10.1016/j.biochi.2008.09.007",
language = "English",
volume = "91",
pages = "252--60",
journal = "BIOCHIMIE",
issn = "0300-9084",
publisher = "Elsevier",
number = "2",

}

RIS

TY - JOUR

T1 - Regulation of the biochemical function of motif VI of HCV NTPase/helicase by the conserved Phe-loop

AU - Hartjen, Philip

AU - Medom, Berthe Kamdem

AU - Reinholz, Michael

AU - Borowski, Peter

AU - Baier, Andrea

PY - 2009/2

Y1 - 2009/2

N2 - In previous works, we demonstrated a potent inhibition of diverse protein kinase C (PKC) functions by a fragment of nonstructural protein 3 (NS3) of hepatitis C virus (HCV), mainly mediated by the Arg-rich amino acid motif HCV(1487-1500). This sequence is localized on the surface of Domain 2 of the NS3 NTPase/helicase in direct vicinity to a flexible loop that is localized between Val1458 and Thr1476. Here, we assessed the regulation of the accessibility of the Arg-rich amino acid motif for PKC by this loop, using two variants of domain 2. The first construct, termed NS3d2Delta, comprises the complete domain, HCV(1361-1503), devoid the loop. The second variant, NS3d2wt corresponds to wild type domain 2. The results indicated an enhanced inhibitory potential of NS3d2Delta towards rat brain PKC and towards the majority of PKC isoforms. This effect and the accompanying change of the mode of inhibition from a mixed mode, exerted by NS3d2wt to a competitive mode, exerted by NS3d2Delta are caused by the deletion of the loop. Accordingly, the presence of the intact loop abolished the binding of domain 2 to the tailed duplex RNA used as helicase substrate, without affecting the binding of dsDNA. Furthermore, a direct competition of dsRNA and PKC for the same binding site HCV(1487-1500), could be documented. The binding of dsRNA to NS3d2Delta previously overlaid with PPKCalpha was reduced to 30% and completely abolished in case of NS3d2Delta overlaid with cAMP-dependent protein kinase A (PKA).

AB - In previous works, we demonstrated a potent inhibition of diverse protein kinase C (PKC) functions by a fragment of nonstructural protein 3 (NS3) of hepatitis C virus (HCV), mainly mediated by the Arg-rich amino acid motif HCV(1487-1500). This sequence is localized on the surface of Domain 2 of the NS3 NTPase/helicase in direct vicinity to a flexible loop that is localized between Val1458 and Thr1476. Here, we assessed the regulation of the accessibility of the Arg-rich amino acid motif for PKC by this loop, using two variants of domain 2. The first construct, termed NS3d2Delta, comprises the complete domain, HCV(1361-1503), devoid the loop. The second variant, NS3d2wt corresponds to wild type domain 2. The results indicated an enhanced inhibitory potential of NS3d2Delta towards rat brain PKC and towards the majority of PKC isoforms. This effect and the accompanying change of the mode of inhibition from a mixed mode, exerted by NS3d2wt to a competitive mode, exerted by NS3d2Delta are caused by the deletion of the loop. Accordingly, the presence of the intact loop abolished the binding of domain 2 to the tailed duplex RNA used as helicase substrate, without affecting the binding of dsDNA. Furthermore, a direct competition of dsRNA and PKC for the same binding site HCV(1487-1500), could be documented. The binding of dsRNA to NS3d2Delta previously overlaid with PPKCalpha was reduced to 30% and completely abolished in case of NS3d2Delta overlaid with cAMP-dependent protein kinase A (PKA).

KW - Amino Acid Motifs

KW - Amino Acid Sequence

KW - Animals

KW - Binding Sites

KW - Conserved Sequence

KW - Escherichia coli

KW - Hepacivirus

KW - Models, Molecular

KW - Molecular Sequence Data

KW - Nucleoside-Triphosphatase

KW - Plasmids

KW - Protein Isoforms

KW - Protein Kinase C

KW - Protein Kinase C-alpha

KW - Protein Structure, Secondary

KW - Protein Structure, Tertiary

KW - RNA Helicases

KW - Rats

KW - Recombinant Proteins

KW - Sequence Homology, Amino Acid

KW - Viral Nonstructural Proteins

U2 - 10.1016/j.biochi.2008.09.007

DO - 10.1016/j.biochi.2008.09.007

M3 - SCORING: Journal article

C2 - 18951948

VL - 91

SP - 252

EP - 260

JO - BIOCHIMIE

JF - BIOCHIMIE

SN - 0300-9084

IS - 2

ER -