the 3′-terminal hexamer sequence of classical swine fever virus rna plays a role in negatively regulating the ires-mediated translation3u2032末端六聚体的古典猪瘟病毒rna序列在负调节ires-mediated翻译过程中发挥作用.pdfVIP

The 3 9-Terminal Hexamer Sequence of Classical swine fever virus RNA Plays a Role in Negatively Regulating the IRES-Mediated Translation 1 1 2 3 1,4 Shih-Wei Huang , Meng-Yu Chan , Wei-Li Hsu , Chin-Cheng Huang , Ching-Hsiu Tsai * 1 Graduate Institute of Biotechnology, National Chung Hsing University, Taichung, Taiwan, 2 Graduate Institute of Microbiology and Public Health, National Chung Hsing University, Taichung, Taiwan, 3 Department of Hog Cholera, Animal Health Research Institute, Council of Agriculture, Taipei, Taiwan, 4 Graduate Institute of Medical Laboratory Science and Biotechnology, China Medical University, Taichung, Taiwan Abstract The 3 9 untranslated region (UTR) is usually involved in the switch of the translation and replication for a positive-sense RNA virus. To understand the 3 9 UTR involved in an internal ribosome entry site (IRES)-mediated translation in Classical swine fever virus (CSFV), we first confirmed the predicted secondary structure (designated as SLI, SLII, SLIII, and SLIV) by enzymatic probing. Using a reporter assay in which the luciferase expression is under the control of CSFV 59 and 39 UTRs, we found that the 39 UTR harbors the positive and negative regulatory elements for translational control. Unlike other stem loops, SLI acts as a repressor for expression of the reporter gene. The negative cis-acting element in SLI is further mapped to the very 39- end hexamer CGGCCC sequence. Further, the CSFV IRES-mediated translation can be enhanced by the heterologous 39-ends such as the poly(A) or the 39 UTR of Hepatitis C virus (HCV). Interestingly, such an enhancement was repressed by flanking this hexamer to the end of poly(A) or HCV 3 9 UTR. After sequence comparison