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- Title
Multiple ETS Family Proteins Regulate PF4 Gene Expression by Binding to the Same ETS Binding Site.
- Authors
Okada, Yoshiaki; Nobori, Haruaki; Shimizu, Mikiko; Watanabe, Miho; Yonekura, Masaaki; Nakai, Tomoko; Kamikawa, Yuko; Wakimura, Atsuko; Funahashi, Nobuaki; Naruse, Hiroki; Watanabe, Ayako; Yamasaki, Daisuke; Fukada, So-ichiro; Yasui, Kazuta; Matsumoto, Kayoko; Sato, Takahiro; Kitajima, Kenji; Nakano, Toru; Aird, William C.; Doi, Takefumi
- Abstract
In previous studies on the mechanism underlying megakaryocyte-specific gene expression, several ETS motifs were found in each megakaryocyte-specific gene promoter. Although these studies suggested that several ETS family proteins regulate megakaryocyte-specific gene expression, only a few ETS family proteins have been identified. Platelet factor 4 (PF4) is a megakaryocyte-specific gene and its promoter includes multiple ETS motifs. We had previously shown that ETS-1 binds to an ETS motif in the PF4 promoter. However, the functions of the other ETS motifs are still unclear. The goal of this study was to investigate a novel functional ETS motif in the PF4 promoter and identify proteins binding to the motif. In electrophoretic mobility shift assays and a chromatin immunoprecipitation assay, FLI-1, ELF-1, and GABP bound to the -51 ETS site. Expression of FLI-1, ELF-1, and GABP activated the PF4 promoter in HepG2 cells. Mutation of a -51 ETS site attenuated FLI- 1-, ELF-1-, and GABP-mediated transactivation of the promoter. siRNA analysis demonstrated that FLI-1, ELF-1, and GABP regulate PF4 gene expression in HEL cells. Among these three proteins, only FLI-1 synergistically activated the promoter with GATA-1. In addition, only FLI-1 expression was increased during megakaryocytic differentiation. Finally, the importance of the -51 ETS site for the activation of the PF4 promoter during physiological megakaryocytic differentiation was confirmed by a novel reporter gene assay using in vitro ES cell differentiation system. Together, these data suggest that FLI- 1, ELF-1, and GABP regulate PF4 gene expression through the -51 ETS site in megakaryocytes and implicate the differentiation stage-specific regulation of PF4 gene expression by multiple ETS factors.
- Subjects
MOLECULAR biology; GENE expression; MEGAKARYOCYTES; RADIOLIGAND assay; TRANSCRIPTION factors; PROMOTERS (Genetics); PROTEIN binding; SMALL interfering RNA
- Publication
PLoS ONE, 2011, Vol 6, Issue 9, p1
- ISSN
1932-6203
- Publication type
Article
- DOI
10.1371/journal.pone.0024837