tailieunhanh - Báo cáo khoa học: Structure of the HIV-1 Rev response element alone and in complex with regulator of virion (Rev) studied by atomic force microscopy

The interaction of multiple HIV-1 regulator of virion (Rev) proteins with the viral RNA target, the Rev response element (RRE), is critical for nuclear export of incompletely spliced and unspliced viral RNA, and for the onset of the late phase in the viral replication cycle. The heterogeneity of the Rev–RRE complex has made it difficult to study using conventional structural methods. | Structure of the HIV-1 Rev response element alone and in complex with regulator of virion Rev studied by atomic force microscopy Jesper Pallesen1 2 Mingdong Dong1 3 f Flemming Besenbacher1 3 and J0rgen Kjems1 2 1 Interdisciplinary Nanoscience Center iNANO University of Aarhus Denmark 2 Department of Molecular Biology University of Aarhus Denmark 3 Department of Physics and Astronomy University of Aarhus Denmark Keywords atomic force microscopy oligomerization Rev RNA RRE Correspondence J. Kjems Department of Molecular Biology University of Aarhus C. F. Mollers alle Build. 1130 DK-8000 Aarhus C Denmark Fax 45 8619 6500 Tel 45 8942 2686 E-mail jk@ Present addresses Columbia University New York City NY USA fHarvard University Cambridge MA USA Received 28 January 2009 revisied 10 May 2009 accepted 3 June 2009 doi The interaction of multiple HIV-1 regulator of virion Rev proteins with the viral RNA target the Rev response element RRE is critical for nuclear export of incompletely spliced and unspliced viral RNA and for the onset of the late phase in the viral replication cycle. The heterogeneity of the Rev-RRE complex has made it difficult to study using conventional structural methods. In the present study atomic force microscopy is applied to directly visualize the tertiary structure of the RRE RNA alone and in complex with Rev proteins. The appearance of the RRE is compatible with the earlier proposed RRE secondary structure in dimensions and overall shape including a stalk and a head interpreted as stem I and stemloops II-V in the secondary structure model respectively. Atomic force microscopy imaging of the Rev-RRE complex revealed an increased height of the structure both in the stalk and head regions which is in accordance with a binding model in which Rev binding to a high affinity site in stem IIB triggers oligomerization of Rev proteins through cooperative binding along stem I in RRE. The present study demonstrates that