PMID- 23043384
OWN - NLM
STAT- MEDLINE
DCOM- 20130225
LR  - 20121009
IS  - 1365-2893 (Electronic)
IS  - 1352-0504 (Linking)
VI  - 19
IP  - 11
DP  - 2012 Nov
TI  - Modulation of replication efficacy of the hepatitis C virus replicon Con1 by 
      site-directed mutagenesis of an NS4B aminoterminal basic leucine zipper.
PG  - 775-83
LID - 10.1111/j.1365-2893.2012.01605.x [doi]
AB  - The hepatitis C virus (HCV) nonstructural protein 4B (NS4B) is assumed to 
      function as a membrane anchor and protein hub for the viral replication complex. 
      The aim of the current work was to modulate HCV replication efficacy in the 
      subgenomic Con1 replicon by mutations of specific sites within the 
      aminoterminal-located basic leucine zipper (bZIP), a candidate motif for 
      protein-protein interactions involving NS4B. Mutational sites and amino acid 
      substitutes were determined by in-silico sequence analyses of the NS4B-bZIP motif 
      in 357 isolates of HCV genotype 1b from the euHCVdB and LosAlamos database and 
      consecutive analysis of conserved physico-chemical properties at bZIP specific 
      positions. Mutants with predicted minor, medium or major reduction of replication 
      efficacy were tested in the pFKI389neo/NS3-3'/ET plasmid replicon model. Four 
      sites (L25, T29, V39 and W43) of crucial importance for bZIP-mediated protein 
      interaction with predicted apolarity of respective amino acid positions were 
      selected for mutational studies. Substitutes with physico-chemical properties 
      matching the predicted requirements either well (T29A), moderately (L25W, V39W), 
      or insufficiently (T29E, W43E) were associated with slightly improved, moderate 
      and marked decreased replication efficacy, respectively. Spontaneous (T29G) and 
      adaptive (A28G, E40G) mutations occurred in the T29E mutation isolate only and 
      were associated with marked reduction of replication efficacy. The bZIP motif 
      region of NS4B is crucial for RNA replication in the subgenomic Con1 replicon 
      system. RNA replication efficacy can be modulated by site-directed mutagenesis at 
      specific bZIP functional sites. New adaptive amino acid mutations were identified 
      within the HCV NS4B protein.
CI  - (c) 2012 Blackwell Publishing Ltd.
FAU - Welker, M-W
AU  - Welker MW
AD  - Klinikum der Johann Wolfgang Goethe-Universitat, Medizinische Klinik, 
      Theodor-Stern-Kai, Frankfurt am Main, Germany.
FAU - Susser, S
AU  - Susser S
FAU - Welsch, C
AU  - Welsch C
FAU - Perner, D
AU  - Perner D
FAU - Fuller, C
AU  - Fuller C
FAU - Kronenberger, B
AU  - Kronenberger B
FAU - Herrmann, E
AU  - Herrmann E
FAU - Zeuzem, S
AU  - Zeuzem S
FAU - Sarrazin, C
AU  - Sarrazin C
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120401
PL  - England
TA  - J Viral Hepat
JT  - Journal of viral hepatitis
JID - 9435672
RN  - 0 (NS4B protein, flavivirus)
RN  - 0 (RNA, Viral)
RN  - 0 (Viral Nonstructural Proteins)
SB  - IM
MH  - Amino Acid Substitution
MH  - Cell Line
MH  - HEK293 Cells
MH  - Hepacivirus/*genetics/physiology
MH  - Humans
MH  - Leucine Zippers
MH  - Mutagenesis, Site-Directed
MH  - Protein Structure, Tertiary
MH  - RNA, Viral/genetics/metabolism
MH  - *Replicon
MH  - Viral Nonstructural Proteins/*chemistry/*genetics/metabolism
MH  - Virus Replication/*genetics
EDAT- 2012/10/10 06:00
MHDA- 2013/02/26 06:00
CRDT- 2012/10/10 06:00
PHST- 2012/10/10 06:00 [entrez]
PHST- 2012/10/10 06:00 [pubmed]
PHST- 2013/02/26 06:00 [medline]
AID - 10.1111/j.1365-2893.2012.01605.x [doi]
PST - ppublish
SO  - J Viral Hepat. 2012 Nov;19(11):775-83. doi: 10.1111/j.1365-2893.2012.01605.x. 
      Epub 2012 Apr 1.