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Searching for nuclear export elements in hepatitis D virus RNA

World Journal of Virology 2013年第3期2卷 123-135页

作者： Natália Freitas Celso Cunha Department of Microbiology Molecular Genetics and Immunology Kansas University Medical Center Rainbow Boulevard Kansa KS 66160 United States Medical Microbiology Unit Center for Malaria and Tropical Diseases Institute of Hygiene and Tropical Medicine Nova University of Lisbon 1349-008 Lisbon Portugal

AIM: To search for the presence of cis elements in hepatitis D virus(HDV) genomic and antigenomic RNA capable of promoting nuclear export. METHODS: We made use of a well characterized chloramphenicol acetyl-transferase reporter system based on plasmid p DM138. Twenty c DNA fragments corresponding to different HDV genomic and antigenomic RNA sequences were inserted in plasmid pD M138, and used in transfection experiments in Huh7 cells. The relative amounts of HDV RNA in nuclear and cytoplasmic fractions were then determined by realtime polymerase chain reaction and Northern blotting. The secondary structure of the RNA sequences that displayed nuclear export ability was further predicted using a web interface. Finally, the sensitivity to leptomycin B was assessed in order to investigate possible cellular pathways involved in HDV RNA nuclear ***: Analysis of genomic RNA sequences did not allow identifying an unequivocal nuclear export element. However, two regions were found to promote the export of reporter m RNAs with efficiency higher than the negative controls albeit lower than the positive control. These regions correspond to nucleotides 266-489 and 584-920, respectively. In addition, when analyzing antigenomic RNA sequences a nuclear export element was found in positions 214-417. Export mediated by the nuclear export element of HDV antigenomic RNA is sensitive to leptomycin B suggesting a possible role of CRM1 in this transport pathway. CONCLUSION: A cis-acting nuclear export element is present in nucleotides 214-417 of HDV antigenomic RNA.

关键词： Hepatitis D virus Genomic RNA antigenomic RNA Nuclear export Nuclear export element

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