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Factor binding and chromatin modification in the promoter of murine Egr1 gene upon induction

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Abstract

The influence of chromatin on immediate-early gene expression has been studied in a model of Egr1 induction in intact mouse cells. ChIP analysis of factor and RNA polymerase binding reveals that the gene is constitutively poised for transcription in nonstimulated cells, but a repressing chromatin structure hampers productive transcription. Stimulation with phorbol esters results in a transient activation, which starts at 5 min and peaks at 30 min. Quantitative mapping of promoter occupancy by the different factors shows for the first time that no direct competition between SP1 and EGR1 occurs. The phosphorylation of ELK1 and CREB, which involves both the cascades of MEK1/2 and p38 kinases, is required for gene expression, which ceases following the binding of NAB1 and NAB2 to the promoter. The changes in histone acetylation and the differential recruitment of histone-modifying complexes further show the role of chromatin in the activation of this immediate-early gene.

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Abbreviations

CBP:

CREB binding protein

ChIP:

Chromatin immunoprecipitation

CRE:

cAMP-response element

CREB:

cAMP-response element-binding protein

ELK:

Ets-like

ERK:

Extracellular signal-regulated kinase

HAT:

Histone acetyltransferase

HDAC:

Histone deacetylase

IE:

Immediate-early

NAB1/2:

NGFI-A binding protein 1/2

S.E:

Standard error

SP1:

Specificity protein 1

SRE:

Serum response element

SRF:

Serum response factor

TPA:

12-O-Tetradecanoylphorbol-13-acetate

TSS:

Transcription start site

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Acknowledgments

We are very indebted to Dr. J. Sandoval for useful comments and to Drs. Medico and Boccacio for the gift of the MLP29 cell line. This work was supported by grants No. BFU2007-63120 and CSD2006-49 from the Ministry of Science and Innovation (Spain), and ACOMP07/085 from the Generalitat Valenciana to G.L.R. G.T. was a recipient of a fellowship (BMC 2001-2868) from the Ministry of Education (Spain).

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  1. The Chromatin Laboratory, Department of Biochemistry and Molecular Biology, University of Valencia, Dr Moliner, 50, 46100, Burjassot, Valencia, Spain

    Gema Tur, Elena I. Georgieva, Andrés Gagete, Gerardo López-Rodas, José L. Rodríguez & Luis Franco

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  1. Gema Tur
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  2. Elena I. Georgieva
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Correspondence to Luis Franco.

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Tur, G., Georgieva, E.I., Gagete, A. et al. Factor binding and chromatin modification in the promoter of murine Egr1 gene upon induction. Cell. Mol. Life Sci. 67, 4065–4077 (2010). https://doi.org/10.1007/s00018-010-0426-3

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