The amino terminal deletion mutants of hepatitis C virus nonstructural protein NS5A function as transcriptional activators in yeast.
To investigate the biological function of hepatitis C virus (HCV)-NS5A, the NS5A was fused at its N-terminus with the DNA binding domain (DBD) of yeast transcriptional activator GAL4 (GAL4-DBD). The GAL4-DBD alone had no transcriptional activation function. However, a mutant of the GAL4-DBD/NS5A fusion protein, in which 129 amino acid residues were deleted from the N-terminus of NS5A, exhibited strong transcriptional activation in yeast cells, bearing the Escherichia coli lacZ reporter gene encoding the beta-galactosidase under the transcriptional control of GAL4 promoter and TATA box. Further mutational analysis of NS5A revealed that the region between the amino acid residues 130 to 352 were critical for optimal level of transactivation. This region includes two acidic domains and one proline-rich region which have been shown to be involved in the function of several transcriptional activators.
Tanimoto
A
A
Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City 66160-7421, USA.
Ide
Y
Y
Arima
N
N
Sasaguri
Y
Y
Padmanabhan
R
R
D015967:Gene Expression Regulation, Viral; D016174:Hepacivirus / Q000235:genetics*; D011993:Recombinant Fusion Proteins; D012441:Saccharomyces cerevisiae; D017384:Sequence Deletion; D013329:Structure-Activity Relationship; D014157:Transcription Factors / Q000737:chemistry*; D015533:Transcriptional Activation; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000378:metabolism; D012324:RNA-Dependent RNA Polymerase
D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase