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Published on: March 28, 2016
Core gene insertion in hepatitis B virus genotype G functions at both the encoded amino acid sequence and RNA
Jing Zhang1, Li Zong1, Yongxiang Wang1
1Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University, Shanghai, China.
Abstract:
Hepatitis B virus genotype G possesses a 36-nucleotide (nt) insertion at the 5' end of core gene, adding 12 residues to core protein. The insertion markedly increased core protein level irrespective of viral genotype, with the effect reproducible using CMV-core gene construct. Here we used such expression constructs and transient transfection experiments in Huh7 cells to identify the structural bases. The insertion is predicted to create a stem-loop structure 14nt downstream of core gene AUG. A + 1 or + 2 frameshift into the 36nt mitigated enhancement of core protein level. Point mutations to disrupt or restore the stem-loop had opposite effects on core protein expression. Shifting the translation initiation site downstream or further upstream of the stem-loop rendered it inhibitory or no longer stimulatory of core protein expression. Therefore, both the reading frame and a properly positioned stem-loop structure contribute to marked increase in core protein expression by the 36-nt insertion.
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