Regulation of neutrophil and eosinophil secondary granule gene express-ion by transcription factors C/EBPε and PU.1
Regulation of neutrophil and eosinophil secondary granule gene express-ion by transcription factors C/EBPε and PU.1
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DOI：
10.1182/blood-2002-04-1039
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发表时间：
2003-04-15
期刊：
影响因子：
20.3
通讯作者：
Koeffler, HP
中科院分区：
文献类型：
--
作者：
Gombart, AF;Kwok, SH;Koeffler, HP
在 C/EBPepsilon(-/-) 小鼠的骨髓中，乳铁蛋白 (LF)、中性粒细胞明胶酶 (NG)、鼠类凯瑟琳样蛋白 (MCLP) 和凯瑟琳 B9 的中性粒细胞二级和三级颗粒 mRNA 不表达；中性粒细胞胶原酶 (NC) 和中性粒细胞明胶酶相关脂质运载蛋白 (NGAL) 严重降低。此外，嗜酸性粒细胞颗粒基因的表达；主要碱性蛋白（MBP）和嗜酸性粒细胞过氧化物酶（EPX）缺失。这些小鼠在骨髓中表达 C/EBPα、C/EBPβ 和 C/EBPδ，其水平与野生型小鼠相似，表明该家族在体内缺乏功能冗余。小鼠成纤维细胞系 NIH 3T3 中 C/EBPepsilon 和 C/EBPα 的稳定诱导表达激活了 B9、MCLP、NC 和 NGAL 的 mRNA 表达，但不激活 LF 的 mRNA 表达。在 C/EBPepsilon 和 C/EBPalpha 的瞬时转染中，B9 被强烈诱导，而其他基因的诱导较弱。 C/EBPbeta 和 C/EBPdelta 蛋白微弱地诱导 B9 表达，但 C/EBPdelta 比其他 C/EBP 更有效地诱导 NC 表达。单独使用C/EBPepsilon 诱导MBP 的表达效率较低，而使用C/EBPepsilon 和GATA-1 诱导MBP 的表达效果较弱，但添加PU.1 导致NIH 3T3 细胞中MBP 的显着协同诱导。人类 MBP 启动子-荧光素酶报告基因构建体中预测的 PU.1 位点的突变消除了对 PU.1 的响应。凝胶迁移分析证明了 PU.1 与该位点的结合。从 PU.1(-/-) 小鼠胚胎肝脏建立的 PU.1 无效骨髓细胞系中不存在 MBP 和 EPX mRNA。 PU.1蛋白表达的恢复恢复了MBP和EPX蛋白表达。这项研究表明，C/EBPepsilon 对于中性粒细胞次级颗粒基因的特定子集的表达是必需的和充分的。此外，它表明PU.1在嗜酸性粒细胞颗粒基因协同激活中的重要性。 (C) 2003 年，美国血液学会。
In the bone marrow of C/EBPepsilon(-/-) mice, expression of neutrophil secondary and tertiary granule mRNAs is absent for lactoferrin (LF), neutrophil gelatinase (NG), murine cathelinlike protein (MCLP), and the cathelin B9; it is severely reduced for neutrophil collagenase (NC) and neutrophil gelatinase-associated lipocalin (NGAL). In addition, the expression of eosinophil granule genes; major basic protein (MBP), and eosinophil peroxidase (EPX) is absent. These mice express C/EBPalpha, C/EBPbeta, and C/EBPdelta in the bone marrow at levels similar to those of their wild-type counterparts, suggesting a lack of functional redundancy among the family in vivo. Stable inducible expression of C/EBPepsilon and C/EBPalpha in the murine fibroblast cell line NIH 3T3 activated expression of mRNAs for B9, MCLP, NC, and NGAL but not for LF. In transient transfections of C/EBPepsilon and C/EBPalpha, B9 was strongly induced with weaker induction of the other genes. C/EBPbeta and C/EBPdelta proteins weakly induced B9 expression, but C/EBPdelta induced NC expression more efficiently than the other C/EBPs. The expression of MBP was inefficiently induced by C/EBPepsilon alone and weakly induced with C/EBPepsilon and GATA-1, but the addition of PU.1 resulted in a striking cooperative induction of MBP in NIH 3T3 cells. Mutation of a predicted PU.1 site in the human MBP promoter-luciferase re porter construct abrogated the response to PU.1. Gel-shift analysis demonstrated binding of PU.1 to this site. MBP and EPX mRNAs were absent in a PU.1-null myeloid cell line established from the embryonic liver of PU.1(-/-) mice. Restitution of PU.1 protein expression restored MBP and EPX protein expression. This study demonstrates that C/EBPepsilon is essential and sufficient for the expression of a particular subset of neutrophil secondary granule genes. Furthermore, it indicates the importance of PU.1 in the cooperative activation of eosinophil granule genes. (C) 2003 by The American Society of Hematology.