DNA bending by transcription factors CREM and CREB.
DNA bending is postulated to be a major determinant of gene expression and has been shown to be specifically induced by some regulatory proteins with DNA-binding properties. Here we show that nuclear factors which naturally bind to CREs (cAMP-responsive elements) are able to induce bending in the sequences flanking this recognition site. In our assays we used a permutated binding site/gel retardation assay and bacterially generated nuclear factors. We have been studying the cAMP-responsive-element modulator (CREM) gene, which encodes both repressors (CREM alpha, beta and gamma) and an activator (CREM tau) of cAMP-responsive transcription by alternative splicing. In addition, two alternative DNA-binding domains can be encoded in different CREM isoforms. No differences in induction of DNA bending by the CREM proteins with the two DNA binding domains were detected. The activator CREB induced DNA bending in a fashion similar to CREM. Importantly, we show that phosphorylation of CREM or CREB alters their mobilities in a regular gel shift assay as well as enhances the angle of DNA bending induced by these proteins.
de Groot
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RP
Laboratoire Génétique Moléculaire des Eucaryotes, CNRS, U184 de l'INSERM, Faculté de Médecine, Institut de Chimie Biologique, Strasbourg, France.
Delmas
V
V
Sassone-Corsi
P
P
D001483:Base Sequence; D051786:Cyclic AMP Response Element Modulator; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism / Q000502:physiology*; D004247:DNA / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D015870:Gene Expression / Q000235:genetics; D007536:Isomerism; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010766:Phosphorylation; D012097:Repressor Proteins; D014158:Transcription, Genetic / Q000235:genetics
D017362:Cyclic AMP Response Element-Binding Protein; D004268:DNA-Binding Proteins; D012097:Repressor Proteins; D051786:Cyclic AMP Response Element Modulator; D004247:DNA