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PMID:19943855

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Citation

Morales-Lázaro, SL, González-Ramírez, R, Gómez, P, Tapia-Ramírez, V, de León, MB and Cisneros, B (2010) Induction of dystrophin Dp71 expression during neuronal differentiation: opposite roles of Sp1 and AP2alpha in Dp71 promoter activity. J. Neurochem. 112:474-85

Abstract

In this study, we delineated the molecular mechanisms that modulate Dp71 expression during neuronal differentiation, using the N1E-115 cell line. We demonstrated that Dp71 expression is up-regulated in response to cAMP-mediated neuronal differentiation of these cells, and that this induction is controlled at promoter level. Functional deletion analysis of the Dp71 promoter revealed that a 5'-flanking 159-bp DNA fragment that contains Sp1 and AP2 binding sites is necessary and sufficient for basal expression of this TATA-less promoter, as well as for its induction during neuronal differentiation. Electrophoretic mobility shift and chromatin immunoprecipitation assays revealed that Sp1 and AP2alpha bind to their respective DNA elements within the Dp71 basal promoter. Overall, mutagenesis assays on the Sp1 and AP2 binding sites, over-expression of Sp1 and AP2alpha, as well as knock-down experiments on Sp1 and AP2alpha gene expression established that Dp71 basal expression is controlled by the combined action of Sp1 and AP2alpha, which act as activator and repressor, respectively. Furthermore, we demonstrated that induction of Dp71 expression in differentiated cells is the result of the maintenance of positive regulation exerted by Sp1, as well as of the loss of AP2alpha binding, which ultimately releases the promoter from repression.

Links

PubMed Online version:10.1111/j.1471-4159.2009.06467.x

Keywords

Animals; Brain/cytology; Bucladesine/pharmacology; Cell Differentiation/drug effects; Cell Differentiation/physiology; Chloramphenicol O-Acetyltransferase/metabolism; Chromatin Immunoprecipitation/methods; DNA-Binding Proteins/metabolism; Dimethyl Sulfoxide/pharmacology; Dystrophin/metabolism; Electrophoretic Mobility Shift Assay/methods; Fatty Acid-Binding Proteins; Free Radical Scavengers/pharmacology; Immunoglobulins/metabolism; Mice; Mutagenesis, Site-Directed/methods; Neuroblastoma; Neurons/drug effects; Neurons/physiology; Nitric Oxide Synthase Type I/metabolism; Promoter Regions, Genetic/drug effects; RNA, Small Interfering/pharmacology; Transfection/methods; Up-Regulation/physiology; beta-Galactosidase/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

MOUSE:SP3

enables

GO:0000987: cis-regulatory region sequence-specific DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

MOUSE:SP1

enables

GO:0000987: cis-regulatory region sequence-specific DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

MOUSE:SP1

enables

GO:0001228: DNA-binding transcription activator activity, RNA polymerase II-specific

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

MOUSE:AP2A

enables

GO:0000987: cis-regulatory region sequence-specific DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

MOUSE:AP2A

enables

GO:0001227: DNA-binding transcription repressor activity, RNA polymerase II-specific

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

See also

References

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