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Molecular and Cellular Biology, October 2000, p. 7247-7258, Vol. 20, No. 19
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

dCtBP-Dependent and -Independent Repression Activities of the Drosophila Knirps Protein

Scott A. Keller,1 Yifan Mao,1,dagger Paolo Struffi,1 Carla Margulies,1,Dagger Catherine E. Yurk,1 Amelia R. Anderson,1 Roxane L. Amey,1 Sarah Moore,1 Julie M. Ebels,1 Kathy Foley,1 Maria Corado,2 and David N. Arnosti1,*

Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, Michigan 48824-1319,1 and Department of Biology, New York University, New York, New York 100032

Received 23 May 2000/Accepted 12 June 2000

Transcriptional repressor proteins play essential roles in controlling the correct temporal and spatial patterns of gene expression in Drosophila melanogaster embryogenesis. Repressors such as Knirps, Krüppel, and Snail mediate short-range repression and interact with the dCtBP corepressor. The mechanism by which short-range repressors block transcription is not well understood; therefore, we have undertaken a detailed structure-function analysis of the Knirps protein. To provide a physiological setting for measurement of repression, the activities of endogenous or chimeric Knirps repressor proteins were assayed on integrated reporter genes in transgenic embryos. Two distinct repression functions were identified in Knirps. One repression activity depends on dCtBP binding, and this function maps to a C-terminal region of Knirps that contains a dCtBP binding motif. In addition, an N-terminal region was identified that represses in a CtBP mutant background and does not bind to the dCtBP protein in vitro. Although the dCtBP protein is important for Knirps activity on some genes, one endogenous target of the Knirps protein, the even-skipped stripe 3 enhancer, is not derepressed in a CtBP mutant. These results indicate that Knirps can utilize two different pathways to mediate transcriptional repression and suggest that the phenomenon of short-range repression may be a combination of independent activities.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI 48824-1319. Phone: (517) 432-5504. Fax: (517) 353-9334. E-mail: arnosti{at}pilot.msu.edu.

dagger Present address: Calydon, Sunnyvale, CA 94089.

Dagger Present address: Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724.

Molecular and Cellular Biology, October 2000, p. 7247-7258, Vol. 20, No. 19
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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