Interaction of hepatitis C virus NS5A with La protein revealed by T7 phage display.
Although the hepatitis C virus (HCV) genome is synthesized by the virus-encoded RNA-dependent RNA polymerase NS5B, other viral and cellular factors are assumed to be required for template-specific initiation and regulation of RNA-synthesis. The cellular protein La, which normally associates with RNA polymerase III transcripts, also interacts with the 5'- and 3'-untranslated regions of several RNA viruses, including HCV. To investigate whether other viral gene products may be involved in this interaction, we constructed an HCV cDNA expression library in bacteriophage T7 allowing portions of the HCV polyprotein to be displayed on the phage surface. Screening of the phage library against La resulted in selection of clones displaying the N-terminal region of HCV NS5A. Co-precipitation of full-length and truncated forms of recombinant NS5A with La revealed that the N-terminal region of NS5A was both necessary and sufficient for binding to La. Although this region of NS5A is essential for HCV replication, the role of the NS5A-La interaction in the infected cell remains to be established.
Houshmand
Hamid
H
Department of Medical Biochemistry and Microbiology, Uppsala University, Biomedical Centre, Uppsala, Sweden.
Bergqvist
Anders
A
D000595:Amino Acid Sequence; D001324:Autoantigens; D017123:Bacteriophage T7 / Q000235:genetics; D018076:DNA, Complementary; D016174:Hepacivirus / Q000235:genetics; D008969:Molecular Sequence Data; D019151:Peptide Library; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012261:Ribonucleoproteins / Q000378:metabolism*; D016415:Sequence Alignment; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*; D000097585:SS-B Antigen; D012324:RNA-Dependent RNA Polymerase
D001324:Autoantigens; D018076:DNA, Complementary; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D012261:Ribonucleoproteins; D017361:Viral Nonstructural Proteins; D000097585:SS-B Antigen; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase