[Transcriptional activation function of hepatitis B virus Pre S1 protein in yeast].
BACKGROUND
To explore the feasibility of cloning of the hepatocyte receptor interacting with the Pre S1 protein of HBV by two hybrid system.


METHODS
Yeast expression plasmids encoding fusion proteins of full length or portions of Pre S1 of HBV and DNA binding domain of yeast protein GAL4 were constructed and used to transform yeast reporter strain SFY526. Reporter gene product ?galactosidase activity was assayed as a measure of transcription activation in yeast. Mammalian expression plasmid encoding fusion proteins of full length Pre S1 and DNA binding domain of GAL4 was constructed and used to cotransfect hepatoma cell line Huh?7 together with CAT reporter plasmid. Cell extracts were assayed for CAT activity by thin?layer chromatography.


RESULTS
The fusion proteins of full length Pre S1 protein and GAL4 DNA binding domain present transcriptional activation function in yeast. The transcription activating sequence is localized to the 21 to 47 amino acids of Pre S1 protein Fusion proteins of full length Pre S 1 and GAL 4 DNA binding domain do not show transcriptional activation function in mammalian cells.


CONCLUSIONS
The transcriptional activating sequence of HBVPre S1 protein in yeast overlaps the hepatocyte receptor binding site. The transcriptional activation function of HBV Pre S1 protein in yeast may prevent researchers?from using yeast two hybrid system to clone HBV receptor interacting with Pre S1 protein. However, the Pre S1 protein does not show transcriptional activation function in mammalian cells. Mammalian two?hybrid system may be a practical method to clone the HBV hepatocyte receptor interacting with Pre S1 protein.
Laboratory of Molecular Biology, Institute of Dermatology, The Second Hospital, Xi?an Jiaotong University, Xi'an 710004, China.
Shengxiang
S
Xiao
Yonglie
Y
Chu
Xuanxian
X
Peng
Cuiling
C
Wang
Shengbin
S
Xiao
Yanhong
Y
Wu
Zhenping
Z
Cao
D004268:DNA-Binding Proteins / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics*; D006514:Hepatitis B Surface Antigens / Q000235:genetics*; D006801:Humans; D011498:Protein Precursors / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D015533:Transcriptional Activation; D014407:Tumor Cells, Cultured; D015003:Yeasts
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C050806:presurface protein 1, hepatitis B surface antigen