RRC ID 56681
Author Yashiro T, Kasakura K, Oda Y, Kitamura N, Inoue A, Nakamura S, Yokoyama H, Fukuyama K, Hara M, Ogawa H, Okumura K, Nishiyama M, Nishiyama C.
Title The hematopoietic cell-specific transcription factor PU.1 is critical for expression of CD11c.
Journal Int Immunol
Abstract PU.1 is a hematopoietic cell-specific transcription factor belonging to the Ets family, which plays an important role in the development of dendritic cells (DCs). CD11c (encoded by Itgax) is well established as a characteristic marker of hematopoietic lineages including DCs. In the present study, we analyzed the role of PU.1 (encoded by Spi-1) in the expression of CD11c. When small interfering RNA (siRNA) for Spi-1 was introduced into bone marrow-derived DCs (BMDCs), the mRNA level and cell surface expression of CD11c were dramatically reduced. Using reporter assays, the TTCC sequence at -56/-53 was identified to be critical for PU.1-mediated activation of the promoter. An EMSA showed that PU.1 directly bound to this region. ChIP assays demonstrated that a significant amount of PU.1 bound to this region on chromosomal DNA in BMDCs, which was decreased in LPS-stimulated BMDCs in accordance with the reduced levels of mRNAs of Itgax and Spi-1, and the histone acetylation degree. Enforced expression of exogenous PU.1 induced the expression of the CD11c protein on the cell surface of mast cells, whereas control transfectants rarely expressed CD11c. Quantitative RT-PCR also showed that the expression of a transcription factor Irf4, which is a partner molecule of PU.1, was reduced in PU.1-knocked down BMDCs. IRF4 transactivated the Itgax gene in a synergistic manner with PU.1. Taken together, these results indicate that PU.1 functions as a positive regulator of CD11c gene expression by directly binding to the Itgax promoter and through transactivation of the Irf4 gene.
Volume 29(2)
Pages 87-94
Published 2017-2-1
DOI 10.1093/intimm/dxx009
PII 3045847
PMID 28338898
MeSH Acetylation Animals CD11c Antigen / genetics CD11c Antigen / metabolism* Cells, Cultured DNA-Binding Proteins / genetics Dendritic Cells / physiology* Gene Expression Regulation Hematopoiesis* / genetics Histones / metabolism Interferon Regulatory Factors / genetics Interferon Regulatory Factors / metabolism* Mice Mice, Inbred BALB C Organ Specificity Promoter Regions, Genetic / genetics Proto-Oncogene Proteins / genetics Proto-Oncogene Proteins / metabolism* RNA, Small Interfering / genetics Trans-Activators / genetics Trans-Activators / metabolism* Transcriptional Activation
IF 4.168
Times Cited 5
Human and Animal Cells CV-1(RCB0160)