PMID- 10544080
OWN - NLM
STAT- MEDLINE
DCOM- 19991126
LR  - 20071114
IS  - 0042-6822 (Print)
IS  - 0042-6822 (Linking)
VI  - 263
IP  - 1
DP  - 1999 Oct 10
TI  - Hepatitis C virus RNA polymerase and NS5A complex with a SNARE-like protein.
PG  - 30-41
AB  - Hepatitis C virus (HCV) NS5A is a phosphoprotein that possesses a cryptic
      trans-activation activity. To investigate its potential role in viral
      replication, we searched for the cellular proteins interacting with NS5A protein 
      by yeast two-hybrid screening of a human hepatocyte cDNA library. We identified a
      newly discovered soluble N-ethylmaleimide-sensitive factor attachment protein
      receptor-like protein termed human vesicle-associated membrane protein-associated
      protein of 33 kDa (hVAP-33). In vitro binding assay and in vivo
      coimmunoprecipitation studies confirmed the interaction between hVAP-33 and NS5A.
      Interestingly, hVAP-33 was also shown to interact with NS5B, the viral
      RNA-dependent RNA polymerase. NS5A and NS5B bind to different domains of hVAP-33:
      NS5A binds to the C-terminus, whereas NS5B binds to the N-terminus of hVAP-33.
      Immunofluorescent staining showed a significant colocalization of hVAP-33 with
      both NS5A and NS5B proteins. hVAP-33 contains a coiled-coil domain followed by a 
      membrane-spanning domain at its C-terminus. Cell fractionation analysis revealed 
      that hVAP-33 is predominantly associated with the ER, the Golgi complex, and the 
      prelysosomal membrane, consistent with its potential role in intracellular
      membrane trafficking. These interactions provide a mechanism for membrane
      association of the HCV RNA replication complex and further suggest that NS5A is a
      part of the viral RNA replication complex.
CI  - Copyright 1999 Academic Press.
FAU - Tu, H
AU  - Tu H
AD  - Department of Molecular Microbiology and Immunology, University of Southern
      California School of Medicine, Los Angeles, California 90033, USA.
FAU - Gao, L
AU  - Gao L
FAU - Shi, S T
AU  - Shi ST
FAU - Taylor, D R
AU  - Taylor DR
FAU - Yang, T
AU  - Yang T
FAU - Mircheff, A K
AU  - Mircheff AK
FAU - Wen, Y
AU  - Wen Y
FAU - Gorbalenya, A E
AU  - Gorbalenya AE
FAU - Hwang, S B
AU  - Hwang SB
FAU - Lai, M M
AU  - Lai MM
LA  - eng
GR  - AI40038/AI/NIAID NIH HHS/United States
GR  - N01-CO-56000/CO/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Virology
JT  - Virology
JID - 0110674
RN  - 0 (Carrier Proteins)
RN  - 0 (Membrane Proteins)
RN  - 0 (NS-5 protein, hepatitis C virus)
RN  - 0 (VAPA protein, human)
RN  - 0 (Vapa protein, rat)
RN  - 0 (Vesicular Transport Proteins)
RN  - 0 (Viral Nonstructural Proteins)
RN  - EC 2.7.7.48 (RNA Replicase)
SB  - IM
MH  - Animals
MH  - COS Cells
MH  - Carrier Proteins/genetics/*metabolism
MH  - Female
MH  - Fluorescent Antibody Technique
MH  - Gene Library
MH  - Hepacivirus/enzymology/genetics/*metabolism/physiology
MH  - Humans
MH  - Membrane Proteins/genetics/*metabolism
MH  - Precipitin Tests
MH  - RNA Replicase/genetics/*metabolism
MH  - Rabbits
MH  - Rats
MH  - Subcellular Fractions
MH  - Two-Hybrid System Techniques
MH  - *Vesicular Transport Proteins
MH  - Viral Nonstructural Proteins/genetics/*metabolism
MH  - Virus Replication
EDAT- 1999/11/02 00:00
MHDA- 1999/11/02 00:01
CRDT- 1999/11/02 00:00
PHST- 1999/11/02 00:00 [pubmed]
PHST- 1999/11/02 00:01 [medline]
PHST- 1999/11/02 00:00 [entrez]
AID - 10.1006/viro.1999.9893 [doi]
AID - S0042-6822(99)99893-8 [pii]
PST - ppublish
SO  - Virology. 1999 Oct 10;263(1):30-41. doi: 10.1006/viro.1999.9893.