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Nucleic Acids Research, 1994, Vol. 22, No. 20 4284-4290
© 1994

STRUCTURAL BIOLOGY

LFB1/HNF1 acts as a repressor of its own transcription

Giulia Piaggio*, Licia Tomei1, Carlo Toniatti1, Raffaele De Francesco1, Jutta Gerstner2,$ and Riccardo Cortese1

Istituto ‘Regina Elena’ Centra Ricerca Sperimentale, Laboratorio Oncogenesi Molecolare via Delle Messi d'Oro 156, 00158 Rome 1IRBM— Istituto di Ricerche di Biologia Molecolare P.Angeletti via Pontina Km 40.600, 00040 Pomezia, Rome, Italy 2EMBL, European Molecular Biology Laboratory MeyerhofstraBe 1, 6200 Heidelberg, Germany

*To whom correspondence should be addressed

Received May 27, 1994. Revised September 5, 1994. Accepted September 5, 1994.

LFB1/HNF1 is a hepatocyte-enriched frans-activator involved in the regulation of many liver-specific genes. We report the cloning and characterization of a rat genomic DNA fragment containing about 3.5 kb of the LFB1/HNF1 gene 5'-flanking region. This DNA segment is capable of directing the liver-specific expression of a reporter gene in transfection assays. More interestingly, the basal activity of the LFB1/HNF1 promoter in cultured hepatoma cell lines is downregulated by exogenously added LFB1/HNF1 protein itself. The ability to repress transcription starting from its own promoter requires the integrity of the N-terminal LFB1/HNF1 DNA-binding domain. Contrary to the expectations, in vitro binding experiments failed to demonstrate any specific and functional interaction of purified LFB1/HNF1 with the -3.5 kb promoter sequence. In addition to the DNA-binding domain, a 60 aa region contained in the C-terminus of the protein and distinct from the previously characterized activation domains, is also required for the repressing function.

$Present address: Leica Vertrieb GmbH, Lilienthalstrafle 39-45, D-6140 Benshcim, Germany


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