p300/cAMP-responsive Element-binding Protein Interactions with Ets-1 and Ets-2 in the Transcriptional Activation of the Human Stromelysin Promoter

Abstract

In this paper we show that transcription factors Ets-1 and Ets-2 recruit transcription adapter proteins p300 and CBP (cAMP-responsive element-binding protein) during the transcriptional activation of the human stromelysin promoter, which contains palindromic Ets-binding sites. Ets-2 and p300/CBP exist as a complex in vivo. Two regions of p300/CBP between amino acids (a.a.) 328 and 596 and a. a. 1678 and 2370 independently can interact with Ets-1 and Ets-2 in vitro and in vivo. Both these regions of p300/CBP bind to the transactivation domain of Ets-2, whereas the C-terminal region binds only to the DNA binding domain of Ets-2. The N- and the C-terminal regions of CBP (a.a. 1-1097 and 1678-2442, respectively) which lack histone acetylation activity independently are capable of coactivating Ets-2. Other Ets family transcription factors failed to cooperate with p300/CBP in stimulating the stromelysin promoter. The LXXLL sequence, reported to be important in receptor-coactivator interactions, does not appear to play a role in the interaction of Ets-2 with p300/CBP. Previous studies have shown that the stimulation of transcriptional activation activity of Ets-2 requires phosphorylation of threonine 72 by the Ras/mitogen-activated protein kinase signaling pathway. We show that mutation of this site does not affect its capacity to bind to and to cooperate with p300/CBP.

Ets-2 and p160 proteins collaborate to regulate c-Myc in endocrine resistant breast cancer

“…Ets-1, -2 and the PEA3 family member, ER81 have been shown to recruit the transcription adaptor proteins p300 and CBP (Yang et al, 1998;Jayaraman et al, 1999;Papoutsopoulou and Janknecht, 2000;Foulds et al, 2004). More recently we have proposed that the steroid receptor coactivator, p160 protein, SRC-1 is a functional coactivator of Ets-2.…”

IL-2 and long-term T cell activation induce physical and functional interaction between STAT5 and ETS transcription factors in human T cells

“…Ets-1 binds various transcription factors, including Sp1 (Gegonne et al, 1993), PEBP2a/AML-1 (Giese et al, 1995), GHF-1/Pit-1 (Bradford et al, 1995), MafB (Sieweke et al, 1996), NF-kB, AP-1 (Thomas et al, 1997), and USF-1 (Sieweke et al, 1998). Despite its high degree of structural similarity with Ets-1, only one protein-protein interaction between Ets-2 and p300/CBP (Jayaraman et al, 1999) and two transcriptional cooperations with GATA3 (Blumenthal et al, 1999) or ATF/Jun family members (Cirillo et al, 1999) have been reported. Two Stat-Ets cooperations have been described.…”

p300 provides a corepressor function by cooperating with YY1 and HDAC3 to repress c-Myc

“…Promoter reporter assays, in vitro and in vivo protein-binding assays, EMSA and ChIP assays Procedures for GST-pull down assays and EMSA were as described (Jayaraman et al, 1999). For detecting the multiprotein complexes, nuclear extracts were prepared from SaOS2 cells after transfecting with expression plasmids expressing HA-p300, FLAG-YY1 and His-HDAC3 were incubated with an YY1 probe and in the presence of appropriate antibodies for 20 min.…”