MilliporeSigma
  • Home
  • Search Results
  • Hepatitis C virus NS3 protein can activate the Notch-signaling pathway through binding to a transcription factor, SRCAP.

Hepatitis C virus NS3 protein can activate the Notch-signaling pathway through binding to a transcription factor, SRCAP.

PloS one (2011-06-16)
Atsushi Iwai, Tsutomu Takegami, Takuya Shiozaki, Tadaaki Miyazaki
ABSTRACT

Persistent infections of hepatitis C virus (HCV) are known to be a major risk factor for causing hepatocellular carcinomas. Nonstructural protein 3 (NS3) of HCV has serine protease and RNA helicase domains, and is essential for the viral replication. Further, NS3 is also considered to be involved in the development of HCV-induced hepatocellular carcinomas. In this report, we focus on the function of NS3 protein, and propose a novel possible molecular mechanism which is thought to be related to the tumorigenesis caused by the persistent infection of HCV. We identified SRCAP (Snf2-related CBP activator protein) as a NS3 binding protein using yeast two-hybrid screening, and a co-immunoprecipitation assay demonstrated that NS3 can bind to SRCAP in mammalian cells. The results of a reporter gene assay using Hes-1 promoter which is known to be a target gene activated by Notch, indicate that NS3 and SRCAP cooperatively activate the Hes-1 promoter in Hep3B cells. In addition, we show in this report that also p400, which is known as a protein closely resembling SRCAP, would be targeted by NS3. NS3 exhibited binding activity also to the 1449-1808 region of p400 by a co-immunoprecipitation assay, and further the activation of the Notch-mediated transcription of Hes-1 promoter by NS3 decreased significantly by the combined silencing of SRCAP and p400 mRNA using short hairpin RNA. These results suggest that the HCV NS3 protein is involved in the activation of the Notch-signaling pathway through the targeting to both SRCAP and p400.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
StableCell RPMI-1640, With stable glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
RPMI-1640 Medium, With L-glutamine and sodium bicarbonate. Without arginine, leucine, lysine, and phenol red, liquid, sterile-filtered, suitable for cell culture, designed for isotope labeling for cell culture applications
Sigma-Aldrich
RPMI-1640 Medium, With L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
SAFC
RPMI-1640 Medium, Hybri-Max, Modified, with L-glutamine, 4500 mg/L glucose and 15mM HEPES, without sodium bicarbonate, powder, suitable for hybridoma
Sigma-Aldrich
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, clone M2, purified immunoglobulin (Purified IgG1 subclass), buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)
Sigma-Aldrich
RPMI-1640 Medium, Modified, with L-glutamine, without phenol red and sodium bicarbonate, powder, suitable for cell culture
SAFC
RPMI-1640 Medium, HEPES Modification, With 25 mM HEPES, without L-glutamine., liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
RPMI-1640 Medium, Modified, with sodium bicarbonate, without L-glutamine and phenol red, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
RPMI-1640 Medium, With sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
RPMI-1640 Medium, With L-glutamine, without sodium bicarbonate, powder, suitable for cell culture
SAFC
RPMI-1640 Medium, HEPES Modification, with L-glutamine and 25mM HEPES, without sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
RPMI-1640 Medium, Modified, with sodium bicarbonate, without methionine, cystine and L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
RPMI-1640 Medium, Modified, with 20 mM HEPES and L-glutamine, without sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
RPMI-1640 Medium, With L-glutamine, without glucose and sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
RPMI-1640 Medium, Dutch Modification, with sodium bicarbonate and 20mM HEPES, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
SAFC
RPMI-1640 Medium, 10 ×, Without L-glutamine, folic acid and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
SAFC
RPMI-1640 Medium, AutoMod, without L-glutamine and sodium bicarbonate, powder, suitable for cell culture
SAFC
RPMI-1640 Medium, with 2.05 mM L-glutamine, with 25mM HEPES, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
CelLytic NuCLEAR Extraction Kit, For mammalian tissue or cultured cells
Roche
X-tremeGENE HP DNA Transfection Reagent, High-performance polymer reagent for transfecting many cell lines