Nucleic Acids Research, Vol 24, Issue 12 2352-2359, Copyright © 1996 by Oxford University Press
DS Peabody and F Lim
The coat protein of bacteriophage MS2 functions as a symmetric dimer to
bind an asymmetric RNA hairpin. This implies the existence of two
equivalent RNA binding sites related to one another by a 2-fold symmetry
axis. In this view the symmetric binding site defined by mutations
conferring the repressor-defective phenotype is a composite picture of
these two asymmetric sites. In order to determine whether the RNA ligand
interacts with amino acid residues on both subunits of the dimer and in the
hope of constructing a functional map of the RNA binding site, we performed
heterodimer complementation experiments. Taking advantage of the physical
proximity of their N- and C-termini, the two subunits of the dimer were
genetically fused, producing a duplicated coat protein which folds normally
and allows the construction of the functional equivalent of obligatory
heterodimers containing all possible pairwise combinations of the
repressor- defective mutations. The restoration of repressor function in
certain heterodimers shows that a single RNA molecule interacts with both
subunits of the dimer and allows the construction of a functional map of
the binding site.
ARTICLES
Complementation of RNA binding site mutations in MS2 coat protein heterodimers
Department of Cell Biology, University of New Mexico School of Medicine and Cancer Research and Treatment Center, Albuquerque, NM 87131, USA.
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