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Nucleic Acids Research, Vol 24, Issue 5 898-906, Copyright © 1996 by Oxford University Press
WF Shen, CP Chang, S Rozenfeld, G Sauvageau, RK Humphries, M Lu, HJ Lawrence, ML Cleary and C Largman
Eight of the nine homeobox genes of the Hoxb locus encode proteins which
contain a conserved hexapeptide motif upstream from the homeodomain. All
eight proteins (Hoxb-1-Hoxb-8) bind to a target oligonucleotide in the
presence of Pbx1a under conditions where minimal or no binding is detected
for the Hox or Pbx1a proteins alone. The stabilities of the Hox-Pbx1a-DNA
complexes vary >100-fold, with the proteins from the middle of the locus
(Hoxb-5 and Hoxb-6) forming very stable complexes, while Hoxb-4, Hoxb-7 and
Hoxb-8 form complexes of intermediate stability and proteins at the 3'-side
of the locus (Hoxb-1- Hoxb-3) form complexes which are very unstable.
Although Hox-b proteins containing longer linker sequences between the
hexapeptide and homeodomains formed unstable complexes, shortening the
linker did not confer complex stability. Homeodomain swapping experiments
revealed that this motif does not independently determine complex
stability. Naturally occurring variations within the hexapeptides of
specific Hox proteins also do not explain complex stability differences.
However, two core amino acids (tryptophan and methionine) which are
absolutely conserved within the hexapeptide domains appear to be required
for complex formation. Removal of N- and C-terminal flanking regions did
not influence complex stability and the members of paralog group 4 (Hoxa-4,
b-4, c-4 and d-4), which share highly conserved hexapeptides, linkers and
homeodomains but different flanking regions, form complexes of similar
stability. These data suggest that the structural features of Hox proteins
which determine Hox-Pbx1a-DNA complex stability reside within the precise
structural relationships between the homeodomain, hexapeptide and linker
regions.
ARTICLES
Hox homeodomain proteins exhibit selective complex stabilities with Pbx and DNA
Department of Medicine San Francisco Veterans Affairs Medical Center, CA 94121, USA.
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