The ORF3 Protein of Hepatitis E Virus Binds to Src Homology 3 Domains and Activates MAPK

Hasan Korkaya, Shahid Jameel, Dinesh Gupta, Shweta Tyagi, Ravinder Kumar, Mohammad Zafrullah, Manjari Mazumdar, Sunil Kumar Lal, Li Xiaofang, Deepak Sehgal, Suman Ranjan Das, Dinkar Sahal

Research output: Contribution to journalArticle

113 Citations (Scopus)

Abstract

The hepatitis E virus (HEV) is the causative agent of hepatitis E, an acute form of viral hepatitis. The biology and pathogenesis of HEV remain poorly understood. We have used in vitro binding assays to show that the HEV ORF3 protein (pORF3) binds to a number of cellular signal transduction pathway proteins. This includes the protein tyrosine kinases Src, Hck, and Fyn, the p85α regulatory subunit of phosphatidylinositol 3-kinase, phospholipase Cγ, and the adaptor protein Grb2. A yeast two-hybrid assay was used to further confirm the pORF3-Grb2 interaction. The binding involves a proline-rich region in pORF3 and the src homology 3 (SH3) domains in the cellular proteins. Competition assays and computer-assisted modeling was used to evaluate the binding surfaces and interaction energies of the pORF3-SH3 complex. In pORF3-expressing cells, pp60src was found to associate with an 80-kDa protein, but no activation of the Src kinase was observed in these cells. However, there was increased activity and nuclear localization of ERK in the pORF3-expressing cells. These studies suggest that pORF3 is a viral regulatory protein involved in the modulation of cell signaling. The ORF3 protein of HEV appears to be the first example of a SH3 domain-binding protein encoded by a virus that causes an acute and primarily self-limited infection.

Original languageEnglish (US)
Pages (from-to)42389-42400
Number of pages12
JournalJournal of Biological Chemistry
Volume276
Issue number45
DOIs
StatePublished - Nov 9 2001
Externally publishedYes

Fingerprint

src Homology Domains
Viruses
Hepatitis E virus
Assays
src-Family Kinases
Viral Regulatory and Accessory Proteins
Proteins
Phosphatidylinositol 3-Kinase
Hepatitis E
Cell signaling
Signal transduction
Two-Hybrid System Techniques
Type C Phospholipases
Proline
Yeast
Hepatitis
Signal Transduction
Carrier Proteins
Chemical activation
Modulation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

The ORF3 Protein of Hepatitis E Virus Binds to Src Homology 3 Domains and Activates MAPK. / Korkaya, Hasan; Jameel, Shahid; Gupta, Dinesh; Tyagi, Shweta; Kumar, Ravinder; Zafrullah, Mohammad; Mazumdar, Manjari; Lal, Sunil Kumar; Xiaofang, Li; Sehgal, Deepak; Das, Suman Ranjan; Sahal, Dinkar.

In: Journal of Biological Chemistry, Vol. 276, No. 45, 09.11.2001, p. 42389-42400.

Research output: Contribution to journalArticle

Korkaya, H, Jameel, S, Gupta, D, Tyagi, S, Kumar, R, Zafrullah, M, Mazumdar, M, Lal, SK, Xiaofang, L, Sehgal, D, Das, SR & Sahal, D 2001, 'The ORF3 Protein of Hepatitis E Virus Binds to Src Homology 3 Domains and Activates MAPK', Journal of Biological Chemistry, vol. 276, no. 45, pp. 42389-42400. https://doi.org/10.1074/jbc.M101546200
Korkaya, Hasan ; Jameel, Shahid ; Gupta, Dinesh ; Tyagi, Shweta ; Kumar, Ravinder ; Zafrullah, Mohammad ; Mazumdar, Manjari ; Lal, Sunil Kumar ; Xiaofang, Li ; Sehgal, Deepak ; Das, Suman Ranjan ; Sahal, Dinkar. / The ORF3 Protein of Hepatitis E Virus Binds to Src Homology 3 Domains and Activates MAPK. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 45. pp. 42389-42400.
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