Conserved GXXXG-and S/T-like motifs in the transmembrane domains of NS4B protein are required for hepatitis C virus replication

Qingxia Han, Jason Aligo, David Manna, Kerry Belton, Sree V. Chintapalli, Yoojin Hong, Randen L. Patterson, Damian B. van Rossum, Kouacou V. Konan

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)6464-6479
Number of pages16
JournalJournal of Virology
Volume85
Issue number13
DOIs
StatePublished - Jul 2011
Externally publishedYes

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Hepatitis C virus
Virus Replication
virus replication
Hepacivirus
Proteins
proteins
chimerism
Protein Domains
Mutation
mutation
Membrane Proteins
Genome
Dimerization
Mutant Proteins
genome
dimerization
Life Cycle Stages
Viral Load
viral load
membrane proteins

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Conserved GXXXG-and S/T-like motifs in the transmembrane domains of NS4B protein are required for hepatitis C virus replication. / Han, Qingxia; Aligo, Jason; Manna, David; Belton, Kerry; Chintapalli, Sree V.; Hong, Yoojin; Patterson, Randen L.; van Rossum, Damian B.; Konan, Kouacou V.

In: Journal of Virology, Vol. 85, No. 13, 07.2011, p. 6464-6479.

Research output: Contribution to journalArticle

Han, Q, Aligo, J, Manna, D, Belton, K, Chintapalli, SV, Hong, Y, Patterson, RL, van Rossum, DB & Konan, KV 2011, 'Conserved GXXXG-and S/T-like motifs in the transmembrane domains of NS4B protein are required for hepatitis C virus replication', Journal of Virology, vol. 85, no. 13, pp. 6464-6479. https://doi.org/10.1128/JVI.02298-10
Han, Qingxia ; Aligo, Jason ; Manna, David ; Belton, Kerry ; Chintapalli, Sree V. ; Hong, Yoojin ; Patterson, Randen L. ; van Rossum, Damian B. ; Konan, Kouacou V. / Conserved GXXXG-and S/T-like motifs in the transmembrane domains of NS4B protein are required for hepatitis C virus replication. In: Journal of Virology. 2011 ; Vol. 85, No. 13. pp. 6464-6479.
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