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Journal of Molecular Endocrinology (2006) 36, 139-151    DOI: 10.1677/jme.1.01918
© 2006 Society for Endocrinology

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Temporal recruitment of CCAAT/enhancer-binding proteins to early and late adipogenic promoters in vivo

N Salma, H Xiao and A N Imbalzano

Department of Cell Biology, University of Massachusetts Medical School, 55 Lake Avenue North, Worcester, Massachusetts 01655, USA

(Requests for offprints should be addressed to A N Imbalzano; Email: anthony.imbalzano{at}umassmed.edu.)

The CCAAT/enhancer-binding protein (C/EBP) family of transcriptional regulators is critically important for the activation of adipogenic genes during differentiation. The C/EBPß and {delta} isoforms are rapidly induced upon adipocyte differentiation and are responsible for activating the adipogenic regulators C/EBP{alpha} and peroxisome proliferator activated receptor (PPAR){gamma}2, which together activate the majority of genes expressed in differentiating adipocytes. However, mitosis is required following the induction of adipogenesis, and the activation of C/EBP{alpha} and PPAR{gamma}2 gene expression is delayed until cell division is underway. Previous studies have used electromobility shift assays to suggest that this delay is due, at least in part, to a delay between the induction of C/EBPß protein levels and the acquisition of DNA binding capacity by C/EBPß. Here we used in vivo chromatin immunoprecipitation analysis of the C/EBP{alpha}, PPAR{gamma}2, resistin, adiponectin, and leptin promoters to examine the kinetics of C/EBP protein binding to adipogenic genes in differentiating cells. In contrast to prior studies, we determined that C/EBPß and {delta} were bound to endogenous regulatory sequences controlling the expression of these genes within 1–4 h of adipogenic induction. These results indicated that C/EBPß and {delta} bind not only to genes that are induced early in the adipogenic process but also to genes that are induced much later during differentiation, without a delay between induction of C/EBP protein levels and DNA binding by these proteins. We also showed that each of the genes examined undergoes a transition in vivo from early occupancy by C/EBPß and {delta} to occupancy by C/EBP{alpha} at times that correlate with the induction of C/EBP{alpha} protein levels, demonstrating the generality of the transition during adipogenesis and indicating that the binding of specific C/EBP isoforms does not correlate with timing of expression from each gene. We have concluded that C/EBP family members bind to adipogenic genes in vivo in a manner that follows the induction of C/EBP protein synthesis.




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K. R. Badri, Y. Zhou, U. Dhru, S. Aramgam, and L. Schuger
Effects of the SANT Domain of Tension-Induced/Inhibited Proteins (TIPs), Novel Partners of the Histone Acetyltransferase p300, on p300 Activity and TIP-6-Induced Adipogenesis
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