title: Negative control glucose dependent mediated by the PreS2 region on the translation efficiency of the reporter Sh-bleomycin gene in Saccharomyces cerevisiae.
abstract: Saccharomyces cerevisiae is able to sense and respond to environmental changes such as the availability of carbon sources. In a previous work, we showed that the expression of the PreS2-S gene of HBV in yeast was negatively regulated at the translational level dependent of glucose. In this study, we show that the S mRNA is detected in the polysomes indicating its active translation, while the PreS2-S mRNA was mainly found in monosomes. Moreover, we used the gene reporter assay based on Zeocin resistance, to better characterize the PreS2 region responsible for this control. Two chimeric genes composed of the N- and C-terminal part of the PreS2 fused to the Sh-bleomycin gene conferring the resistance to Zeocin were expressed in yeast. We found that the strain expressing the N-terminal part of the PreS2 was sensitive to Zeocin on rich medium with 2% glucose. In contrast, the strain harbouring the C-terminal part of the PreS2 fused to the Sh-bleomycin grew on Zeocin, indicating that the Sh-bleomycin mRNA is efficiently translated, subsequently conferring resistance to Zeocin. Our data suggest the establishment of a translational control via the N-terminal part of the PreS2 mediated by the presence of 2% glucose in the media.
authors.affiliation: Laboratory of Biomass Valorisation and Production of Eukaryotic Proteins, Center of Biotechnology of Sfax, University of Sfax, Sfax, Tunisia.
authors.forename: Nadia
authors.initials: N
authors.lastname: Hadiji-Abbes
authors.forename: Istabrak
authors.initials: I
authors.lastname: Borchani-Chabchoub
authors.forename: Ali
authors.initials: A
authors.lastname: Gargouri
authors.forename: Raja
authors.initials: R
authors.lastname: Mokdad-Gargouri
mesh_terms: D001761:Bleomycin / Q000494:pharmacology; D025141:Drug Resistance, Fungal; D015870:Gene Expression; D023061:Gene Order; D017930:Genes, Reporter; D005947:Glucose / Q000378:metabolism*; D006514:Hepatitis B Surface Antigens / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011132:Polyribosomes / Q000378:metabolism; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*
chemical_list: D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; C050807:presurface protein 2, hepatitis B surface antigen; D001761:Bleomycin; C105427:Zeocin; D005947:Glucose
keywords: PreS2; Zeocin resistance; gene reporter; hepatitis B virus; translational control