PMID- 21543474
OWN - NLM
STAT- MEDLINE
DCOM- 20110915
LR  - 20211020
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Print)
IS  - 0022-538X (Linking)
VI  - 85
IP  - 14
DP  - 2011 Jul
TI  - NS4B self-interaction through conserved C-terminal elements is required for the 
      establishment of functional hepatitis C virus replication complexes.
PG  - 6963-76
LID - 10.1128/JVI.00502-11 [doi]
AB  - Hepatitis C virus (HCV) is an important human pathogen, persistently infecting 
      more than 170 million individuals worldwide. Studies of the HCV life cycle have 
      become possible with the development of cell culture systems supporting the 
      replication of viral RNA and the production of infectious virus. However, the 
      exact functions of individual proteins, especially of nonstructural protein 4B 
      (NS4B), remain poorly understood. NS4B triggers the formation of specific, 
      vesicular membrane rearrangements, referred to as membranous webs, which have 
      been reported to represent sites of HCV RNA replication. However, the mechanism 
      of vesicle induction is not known. In this study, a panel of 15 mutants carrying 
      substitutions in the highly conserved NS4B C-terminal domain was generated. Five 
      mutations had only a minor effect on replication, but two of them enhanced 
      assembly and release of infectious virus. Ten mutants were replication defective 
      and used for selection of pseudoreversions. Most of the pseudoreversions also 
      localized to the highly conserved NS4B C-terminal domain and were found to 
      restore replication competence upon insertion into the corresponding primary 
      mutant. Importantly, pseudoreversions restoring replication competence also 
      restored heterotypic NS4B self-interaction, which was disrupted by the primary 
      mutation. Finally, electron microscopy analyses of membrane alterations induced 
      by NS4B mutants revealed striking morphological abnormalities, which were 
      restored to wild-type morphology by the corresponding pseudoreversion. These 
      findings demonstrate the important role of the C-terminal domain in NS4B 
      self-interaction and the formation of functional HCV replication complexes.
FAU - Paul, David
AU  - Paul D
AD  - Department of Infectious Diseases, Molecular Virology, Heidelberg University, Im 
      Neuenheimer Feld 345, 69120 Heidelberg, Germany.
FAU - Romero-Brey, Ines
AU  - Romero-Brey I
FAU - Gouttenoire, Jerome
AU  - Gouttenoire J
FAU - Stoitsova, Savina
AU  - Stoitsova S
FAU - Krijnse-Locker, Jacomine
AU  - Krijnse-Locker J
FAU - Moradpour, Darius
AU  - Moradpour D
FAU - Bartenschlager, Ralf
AU  - Bartenschlager R
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110504
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (DNA Primers)
RN  - 0 (NS4B protein, flavivirus)
RN  - 0 (RNA, Viral)
RN  - 0 (Viral Nonstructural Proteins)
SB  - IM
MH  - Base Sequence
MH  - Blotting, Western
MH  - Cell Line
MH  - Conserved Sequence
MH  - DNA Primers
MH  - Fluorescence Resonance Energy Transfer
MH  - Hepacivirus/genetics/*physiology
MH  - Humans
MH  - Mutation
MH  - RNA, Viral/biosynthesis
MH  - Viral Nonstructural Proteins/chemistry/genetics/*physiology
MH  - *Virus Replication
PMC - PMC3126587
EDAT- 2011/05/06 06:00
MHDA- 2011/09/16 06:00
PMCR- 2012/01/01
CRDT- 2011/05/06 06:00
PHST- 2011/05/06 06:00 [entrez]
PHST- 2011/05/06 06:00 [pubmed]
PHST- 2011/09/16 06:00 [medline]
PHST- 2012/01/01 00:00 [pmc-release]
AID - JVI.00502-11 [pii]
AID - 0502-11 [pii]
AID - 10.1128/JVI.00502-11 [doi]
PST - ppublish
SO  - J Virol. 2011 Jul;85(14):6963-76. doi: 10.1128/JVI.00502-11. Epub 2011 May 4.