Linked Life Data

15037615

Source:http://linkedlifedata.com/resource/pubmed/id/15037615

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
28
pubmed:dateCreated
2004-7-5
pubmed:abstractText
Hepatitis E virus (HEV), a plus-stranded RNA virus contains three open reading frames. Of these, ORF1 encodes the viral nonstructural polyprotein; ORF2 encodes the major capsid protein and ORF3 codes for a phosphoprotein of undefined function. Using the yeast two-hybrid system to screen a human cDNA liver library we have isolated, an N-terminal deleted protein, alpha(1) -microglobulin/bikunin precursor (AMBP) that specifically interacts with the ORF3 protein of HEV. Independently cloned, full-length AMBP was obtained and tested positive for interaction with ORF3 using a variety of in vivo and in vitro techniques. AMBP, a liver-specific precursor protein codes for two different unrelated proteins alpha(1)-microglobulin (alpha(1)m) and bikunin. alpha(1) m individually interacted with ORF3. The above findings were validated by COS-1 cell immunoprecipitation, His(6) pull-down experiments, and co-localization experiments followed by fluorescence resonance energy transfer analysis. Human liver cells showing co-localization of ORF3 with endogenously expressing alpha(1) m showed a distinct disappearance of the protein from the Golgi compartment, suggesting that ORF3 enhances the secretion of alpha(1)m out of the hepatocyte. Using drugs to block the secretory pathway, we showed that alpha m was not degraded in the presence of ORF3. Finally, (1)pulse labeling of alpha(1)m showed that its secretion was expedited out of the liver cell at faster rates in the presence of the ORF3 protein. Hence, ORF3 has a direct biological role in enhancing alpha(1)m export from the hepatocyte.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
9
pubmed:volume
279
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
29308-19
pubmed:dateRevised
2008-6-23
pubmed:meshHeading
pubmed-meshheading:15037615-Alpha-Globulins, pubmed-meshheading:15037615-Animals, pubmed-meshheading:15037615-Brefeldin A, pubmed-meshheading:15037615-COS Cells, pubmed-meshheading:15037615-Cell Line, Tumor, pubmed-meshheading:15037615-Fluorescence Resonance Energy Transfer, pubmed-meshheading:15037615-Gene Library, pubmed-meshheading:15037615-Golgi Apparatus, pubmed-meshheading:15037615-Hepatitis E virus, pubmed-meshheading:15037615-Hepatocytes, pubmed-meshheading:15037615-Humans, pubmed-meshheading:15037615-Liver, pubmed-meshheading:15037615-Membrane Glycoproteins, pubmed-meshheading:15037615-Monensin, pubmed-meshheading:15037615-Open Reading Frames, pubmed-meshheading:15037615-Protein Synthesis Inhibitors, pubmed-meshheading:15037615-Protein Transport, pubmed-meshheading:15037615-Trypsin Inhibitor, Kunitz Soybean, pubmed-meshheading:15037615-Two-Hybrid System Techniques, pubmed-meshheading:15037615-Viral Proteins
pubmed:year
2004
pubmed:articleTitle
The ORF3 protein of hepatitis E virus interacts with liver-specific alpha1-microglobulin and its precursor alpha1-microglobulin/bikunin precursor (AMBP) and expedites their export from the hepatocyte.
pubmed:affiliation
Virology Group, International Centre for Genetic Engineering & Biotechnology, P O Box 10504, Aruna Asaf Ali Rd., New Delhi 110067, India.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't