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Nucleic Acids Research, 1993, Vol. 21, No. 17 3951-3957
© 1993

MOLECULAR BIOLOGY

Definition of the DNA-binding site repertoire for the Drosophila transcription factor SNAIL

Viviane Mauhin, Yves Lutz, Christine Dennefeld and Audrey Alberga*

Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS Unité 184 de Biologie Moléculaire et de Génié Génétique de I'INSERM Institut de Chimie Biologique, Faculté de Médecine 11 rue Humann, 67085 Strasbourg Cedex, France

*To whom correspondance should be addressed

Received June 14, 1993. Accepted July 14, 1993.

The Drosophila gene snail (sna) which encodes a zinc finger protein Is essential for dorsal - ventral pattern formation In the developing embryo. We have defined a repertoire of SNAIL (SNA) binding sites using recomblnant SNA proteins to select specific binding sequences from a pool of random sequence nucleotldes. The bound sequences which were selected by multiple rounds of gel retardation and amplification by the polymerase chain reaction (PCR) were subsequently cloned and sequenced. The consensus sequence, 5'G/A A/t G/A A CAGGTG C/t A C 3', with a highly conserved core of 6 bases, CAGGTG, shares no significant homology with known binding sequences of other Drosophila zinc finger proteins. However, the CAGGTG core Is identical to the core motif of aHLH (helix-loop-helix) binding sites. The strongest SNA binding is obtained with sequences containing this core motif whereas reduced binding Is seen for sequences with canonical CANNTG HLH motifs. Interestingly, SNA binding is detected In the promoter region of the snail gene. Transient expression In co-transfection experiments using a SNA binding element (SBE) linked to a heterologous promoter Indicates that SNA has the ability to function as a transcription activator.


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