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Molecular and Cellular Biology, November 2000, p. 8112-8123, Vol. 20, No. 21
Division of Developmental Biology, Children's
Hospital Research Foundation, Cincinnati, Ohio 45229
Received 11 May 2000/Returned for modification 10 July
2000/Accepted 2 August 2000
We describe experiments to compare the activities of two
Drosophila homeodomain proteins, Bicoid (Bcd) and an
altered-specificity mutant of Fushi tarazu, Ftz(Q50K). Although the
homeodomains of these proteins share a virtually indistinguishable
ability to recognize a consensus Bcd site, only Bcd can activate
transcription from natural enhancer elements when assayed in both yeast
and Drosophila Schneider S2 cells. Our analysis of chimeric
proteins suggests that both the homeodomain of Bcd and sequences
outside the homeodomain contribute to its ability to recognize natural enhancer elements. We further show that, unlike the Bcd homeodomain, the Ftz(Q50K) homeodomain fails to recognize nonconsensus sites found
in natural enhancer elements. The defect of a chimeric protein containing the homeodomain of Ftz(Q50K) in place of that of Bcd can be
preferentially restored by converting the nonconsensus sites in natural
enhancer elements to consensus sites. Our experiments suggest that the
biological specificity of Bcd is determined by combinatorial
contributions of two important mechanisms: the nonconsensus site
recognition function conferred by the homeodomain and the cooperativity
function conferred primarily by sequences outside the homeodomain. A
systematic comparison of different assay methods and enhancer elements
further suggests a fluid nature of the requirements for these two Bcd
functions in target selection.
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Target Selectivity of Bicoid Is Dependent on
Nonconsensus Site Recognition and Protein-Protein Interaction
*
Corresponding author. Mailing address: Division of
Developmental Biology, Children's Hospital Research Foundation, 3333 Burnet Ave., Cincinnati, OH 45229. Phone: (513) 636-7977. Fax: (513) 636-4317. E-mail: jun.ma{at}chmcc.org.
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