PMID- 21507970
OWN - NLM
STAT- MEDLINE
DCOM- 20110825
LR  - 20211020
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Print)
IS  - 0022-538X (Linking)
VI  - 85
IP  - 13
DP  - 2011 Jul
TI  - Conserved GXXXG- and S/T-like motifs in the transmembrane domains of NS4B protein 
      are required for hepatitis C virus replication.
PG  - 6464-79
LID - 10.1128/JVI.02298-10 [doi]
AB  - Hepatitis C virus (HCV) nonstructural protein 4B (NS4B) is an integral membrane 
      protein, which plays an important role in the organization and function of the 
      HCV replication complex (RC). Although much is understood about its amphipathic 
      N-terminal and C-terminal domains, we know very little about the role of the 
      transmembrane domains (TMDs) in NS4B function. We hypothesized that in addition 
      to anchoring NS4B into host membranes, the TMDs are engaged in intra- and 
      intermolecular interactions required for NS4B structure/function. To test this 
      hypothesis, we have engineered a chimeric JFH1 genome containing the Con1 NS4B 
      TMD region. The resulting virus titers were greatly reduced from those of JFH1, 
      and further analysis indicated a defect in genome replication. We have mapped 
      this incompatibility to NS4B TMD1 and TMD2 sequences, and we have defined 
      putative TMD dimerization motifs (GXXXG in TMD2 and TMD3; the S/T cluster in 
      TMD1) as key structural/functional determinants. Mutations in each of the 
      putative motifs led to significant decreases in JFH1 replication. Like most of 
      the NS4B chimeras, mutant proteins had no negative impact on NS4B membrane 
      association. However, some mutations led to disruption of NS4B foci, implying 
      that the TMDs play a role in HCV RC formation. Further examination indicated that 
      the loss of NS4B foci correlates with the destabilization of NS4B protein. 
      Finally, we have identified an adaptive mutation in the NS4B TMD2 sequence that 
      has compensatory effects on JFH1 chimera replication. Taken together, these data 
      underscore the functional importance of NS4B TMDs in the HCV life cycle.
FAU - Han, Qingxia
AU  - Han Q
AD  - Department of Biochemistry and Molecular Biology, The Pennsylvania State 
      University, 308 Althouse Laboratory, University Park, PA 16802, USA.
FAU - Aligo, Jason
AU  - Aligo J
FAU - Manna, David
AU  - Manna D
FAU - Belton, Kerry
AU  - Belton K
FAU - Chintapalli, Sree V
AU  - Chintapalli SV
FAU - Hong, Yoojin
AU  - Hong Y
FAU - Patterson, Randen L
AU  - Patterson RL
FAU - van Rossum, Damian B
AU  - van Rossum DB
FAU - Konan, Kouacou V
AU  - Konan KV
LA  - eng
GR  - K22CA129241/CA/NCI NIH HHS/United States
GR  - R01 GM087410-01/GM/NIGMS NIH HHS/United States
GR  - R25 GM078675/GM/NIGMS NIH HHS/United States
GR  - R01 GM087410/GM/NIGMS NIH HHS/United States
GR  - K22 CA129241/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20110420
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (NS4B protein, flavivirus)
RN  - 0 (RNA, Viral)
RN  - 0 (Viral Nonstructural Proteins)
SB  - IM
MH  - *Amino Acid Motifs
MH  - Amino Acid Sequence
MH  - Cell Line, Tumor
MH  - Cell Membrane/metabolism
MH  - *Consensus Sequence
MH  - Hepacivirus/classification/genetics/*physiology
MH  - Humans
MH  - Molecular Sequence Data
MH  - Mutation
MH  - Protein Multimerization
MH  - RNA, Viral/genetics/metabolism
MH  - Sequence Alignment
MH  - Structure-Activity Relationship
MH  - Viral Nonstructural Proteins/*chemistry/genetics/*metabolism
MH  - *Virus Replication
PMC - PMC3126491
EDAT- 2011/04/22 06:00
MHDA- 2011/08/27 06:00
PMCR- 2012/01/01
CRDT- 2011/04/22 06:00
PHST- 2011/04/22 06:00 [entrez]
PHST- 2011/04/22 06:00 [pubmed]
PHST- 2011/08/27 06:00 [medline]
PHST- 2012/01/01 00:00 [pmc-release]
AID - JVI.02298-10 [pii]
AID - 2298-10 [pii]
AID - 10.1128/JVI.02298-10 [doi]
PST - ppublish
SO  - J Virol. 2011 Jul;85(13):6464-79. doi: 10.1128/JVI.02298-10. Epub 2011 Apr 20.