Phosphorylation and Acetylation of Histone H3 and Autoregulation by Early Growth Response 1 Mediate Interleukin 1β Induction of Early Growth Response 1 Transcription

Abstract

Objective-The transcription factor early growth response (EGR)-1 has been implicated as a key vascular phenotypic switch through its control of inducible transcription. EGR-1 autoregulation, and histone modification in the EGR-1 promoter, represent key mechanisms in EGR-1 control, but have not been explored. Methods and Results-We demonstrate that EGR-1 regulates its own transcription and that this involves histone H3 phosphorylation and acetylation. EGR-1 transactivates its promoter in smooth muscle cells exposed to interleukin (IL) 1␤ through a novel cis-acting element (Ϫ211/Ϫ203). PD98059, which inhibits mitogen-activated protein kinase kinase/extracellular regulated kinase (MEK/ERK) attenuates IL-1␤-inducible phosphorylation of extracellular signalregulated kinase 1/2 and mitogen and stress-activated protein kinases 1/2; and reduces levels of phosphorylated and acetylated histone H3. Histone deacetylase inhibition enhances EGR-1 transcription in response to cytokine. Conversely, suppression of histone modification with mitogen and stress-activated protein kinase 1/2 short interfering RNA, or the histone H3 acetyltransferase inhibitor Garcinol, inhibits IL-1␤-inducible EGR-1 transcription. EGR-1 interacts with the acetyltransferase p300. Acetylated H3 and phosphorylated H3 are enriched at the promoter of EGR-1; and EGR-1 is enriched at the promoters of tissue factor and plasminogen activator inhibitor 1 in response to IL-1␤, and attenuated by PD98059, Garcinol, and mitogen and stress-activated protein kinase 1/2 short interfering RNA. Key Words: histone modification Ⅲ immediate-early gene Ⅲ transcription E arly growth response 1 (human EGR-1 or mouse EGR-1, also known as NGFI-A, Zif268, Krox24, and Tis8) is an inducible zinc finger transcription factor that plays a critical role in controlling cell growth, proliferation, differentiation, and apoptosis. 1-5 EGR-1 belongs to a family of zinc finger transcription factors that also includes EGR-2, EGR-3, EGR-4, EGR-␣, and the Wilm tumor gene product. 6 EGR-1 contains a highly conserved DNA-binding domain (three zinc finger domains of the C2H2 subtype), an activation domain, a repression domain, and a nuclear localization signal. 7,8 Carman and Monroe 9 identified an activation domain between amino acids 174 and 270, a serine/threonine/prolinerich region that is critical for its activity. Mutants lacking this domain perturb wild-type EGR-1 function. 9 EGR-1 regulates the transcription of target genes by binding to GC-rich consensus DNA elements present in the regulatory regions in vascular pathophysiologically relevant genes, such as insulinlike growth factor II, platelet-derived growth factor A and B chains, tissue factor, fibroblast growth factor 2, transforming growth factor ␤1, urokinase plasminogen activator, and plasminogen activator inhibitor (PAI) 1. 5,7,10 -13 EGR-1 also regulates the expression of its corepressor nerve growth factor I-A binding protein-2. 11 Functional cis-acting elements within the human EGR-1 promoter have been identified in differen...

Dimethylcelecoxib inhibits mPGES-1 promoter activity by influencing EGR1 and NF-κB

“…It is known that EGR1 expression is regulated by EGR1 itself, NF-κB, serum response elements and AP-1 [21][22]. Autoregulation of EGR1 is dependent on phosphorylation and acetylation of histone H3 [22].…”

Determining PTEN Functional Status by Network Component Deduced Transcription Factor Activities

“…PTEN -inducible mouse Pten Δloxp/Δloxp MEF cells [21] or PC3 [6], [54] cells were cultured in DMEM or RPMI medium, respectively, supplemented with 10% tetracycline-free fetal calf serum (Invitrogen), 100 U/ml penicillin and streptomycin (Gibco).…”

Histone H3 phosphorylation, immediate-early gene expression, and the nucleosomal response: a historical perspective1This article is part of Special Issue entitled Asilomar Chromatin and has undergone the Journal’s usual peer review process.

“…synthetic H3 peptides reported that the binding affinity increased 10-fold when the peptide was phosphorylated on serine 10 relative to the unmodified peptide (Cheung et al 2000). As well, a number of studies reported a localized increase in H3 acetylation at different promoters regulated by H3S10ph, which was in turn reduced when MSK1/2 was inhibited (Agalioti et al 2002;Cheung et al 2000;Jeong et al 2010;Wang et al 2010). One study mapped the distribution of H3K9ac as well as trimethylation of lysine 4 (H3K4me3), lysine 36 (H3K36me3), and lysine 79 (H3K79me3) at various immediate-early gene promoter and coding regions following induction (Edmunds et al 2008).…”

“…HDAC activity was found to be decreased, leading to an increase in H3 acetylation (H3K14ac). Increased immediate-early gene expression was observed following preincubation with an HDAC inhibitor, resulting in increased levels of both H3 phosphorylation (both H3S10ph and H3S28ph) and acetylation (typically H3K9ac and H3K14ac) (Jeong et al 2010;Wang et al 2010;Zhong et al 2003). However, contrary to expectation, these results are contested by a similar study, which observed inhibited expression of the immediate-early gene Jun with increasing incubation time and concentrations of HDAC inhibitors along with a variety of known stimulants (e.g., TPA and EGF), while acetylated and phosphosphorylated H3 increased as expected (Hazzalin and Mahadevan 2005).…”