Nucleic Acids Research, Vol 24, Issue 15 2919-2923, Copyright © 1996 by Oxford University Press
JS Greatorex, V Laisse, MC Dockhelar and AM Lever
The formation of a genomic RNA dimer appears to be a critical step in the
life cycle of all retroviruses. To investigate the site and nucleotide
interactions involved in this process, a 531 bp DNA fragment encompassing
sequences up- and downstream of the splice donor in human T cell leukaemia
virus type 1 (HTLV-1) was inserted into a plasmid vector under the control
of the SP6 promoter. RNA transcripts generated in vitro from this template
formed dimers which could be dissociated by heating at 60-80 degrees C for
3 min. The physical properties of the dimeric RNA were not consistent with
either Watson-Crick base pairing or guanine tetrad formation as being
solely responsible for the interaction. Deletion mutagenesis identified a
32 nt sequence required for dimerisation. Computer modelling was carried
out in order to identify putative RNA secondary structures within this
essential region. A stem-loop structure was identified, the stem of which
was conserved among different sequenced isolates of HTLV-1. This sequence
also contains a 15 nt palindrome. We sought by disruptive and compensatory
mutagenesis to define the possible roles of these two structures in dimer
linkage.
ARTICLES
Sequences involved in the dimerisation of human T cell leukaemia virus type-1 RNA
Department of Medicine, University of Cambridge, UK.
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