Ribozyme-mediated inhibition of HIV 1 suggests nucleolar trafficking of HIV-1 RNA.
The HIV regulatory proteins Tat and Rev have a nucleolar localization property in human cells. However, no functional role has been attributed to this localization. Recently it has been demonstrated that expression of Rev induces nucleolar relocalization of some protein factors involved in Rev export. Because the function of Rev is to bind HIV RNA and facilitate transport of singly spliced and unspliced RNA to the cytoplasm, it is likely that the nucleolus plays a critical role in HIV-1 RNA export. As a test for trafficking of HIV-1 RNAs into the nucleolus, a hammerhead ribozyme that specifically cleaves HIV-1 RNA was inserted into the body of the U16 small nucleolar RNA, resulting in accumulation of the ribozyme within the nucleoli of human cells. HeLa CD4(+) and T cells expressing this nucleolar localized ribozyme exhibit dramatically suppressed HIV-1 replication. The results presented here suggest a trafficking of HIV-1 RNA through the nucleoli of human cells, thus posing a different paradigm for lentiviral RNA processing.
Department of Molecular Biology, Beckman Research Institute of the City of Hope, and Graduate School of Biological Sciences, City of Hope, Duarte, CA 91010-3011, USA.
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Michienzi
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Cagnon
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Bahner
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Rossi
D001483:Base Sequence; D002460:Cell Line; D002466:Cell Nucleolus / Q000821:virology*; D017931:DNA Primers; D015694:Gene Products, rev / Q000502:physiology; D015696:Gene Products, tat / Q000502:physiology; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008561:Membrane Fusion / Q000502:physiology*; D016337:RNA, Catalytic / Q000502:physiology*; D054321:rev Gene Products, Human Immunodeficiency Virus; D054322:tat Gene Products, Human Immunodeficiency Virus
D017931:DNA Primers; D015694:Gene Products, rev; D015696:Gene Products, tat; D016337:RNA, Catalytic; D054321:rev Gene Products, Human Immunodeficiency Virus; D054322:tat Gene Products, Human Immunodeficiency Virus