Mancl Margo E, Hu Guodong, Sangster-Guity Niquiche, Olshalsky Stacey L, Hoops Katherine, Fitzgerald-Bocarsly Patricia, Pitha Paula M, Pinder Karen, Barnes Betsy J
Division of Viral Oncology, The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21231, USA.
J Biol Chem. 2005 Jun 3;280(22):21078-90. doi: 10.1074/jbc.M500543200. Epub 2005 Mar 31.
Interferon regulatory factor-5 (IRF-5) is a mediator of virus-induced immune activation and type I interferon (IFN) gene regulation. In human primary plasmacytoid dendritic cells (PDC), IRF-5 is transcribed into four distinct alternatively spliced isoforms (V1, V2, V3, and V4), whereas in human primary peripheral blood mononuclear cells two additional new isoforms (V5 and V6) were identified. The IRF-5 V1, V2, and V3 transcripts have different noncoding first exons and distinct insertion/deletion patterns in exon 6. Here we showed that V1 and V3 have distinct transcription start sites and are regulated by two discrete promoters. The V1 promoter (P-V1) is constitutively active, contains an IRF-E consensus-binding site, and is further stimulated in virus-infected cells by IRF family members. In contrast, endogenous V3 transcripts were up-regulated by type I IFNs, and the V3 promoter (P-V3) contains an IFN-stimulated responsive element-binding site that confers responsiveness to IFN through binding of the ISGF3 complex. In addition to V5 and V6, we have identified three more alternatively spliced IRF-5 isoforms (V7, V8, and V9); V5 and V6 were expressed in peripheral blood mononuclear cells from healthy donors and in immortalized B and T cell malignancies, whereas expression of V7, V8, and V9 transcripts were detected only in human cancers. The results of this study demonstrated the existence of multiple IRF-5 spliced isoforms with distinct cell type-specific expression, cellular localization, differential regulation, and dissimilar functions in virus-mediated type I IFN gene induction.
干扰素调节因子5(IRF-5)是病毒诱导的免疫激活和I型干扰素(IFN)基因调控的介质。在人原代浆细胞样树突状细胞(PDC)中,IRF-5转录为四种不同的可变剪接异构体(V1、V2、V3和V4),而在人原代外周血单个核细胞中鉴定出另外两种新的异构体(V5和V6)。IRF-5 V1、V2和V3转录本具有不同的非编码第一外显子,并且在外显子6中有不同的插入/缺失模式。在这里,我们表明V1和V3具有不同的转录起始位点,并受两个离散启动子的调控。V1启动子(P-V1)组成性激活,包含一个IRF-E共有结合位点,并且在病毒感染的细胞中被IRF家族成员进一步刺激。相比之下,内源性V3转录本被I型IFN上调,并且V3启动子(P-V3)包含一个IFN刺激反应元件结合位点,该位点通过ISGF3复合物的结合赋予对IFN的反应性。除了V5和V6,我们还鉴定出另外三种可变剪接的IRF-5异构体(V7、V8和V9);V5和V6在健康供体的外周血单个核细胞以及永生化的B和T细胞恶性肿瘤中表达,而V7、V8和V9转录本仅在人类癌症中检测到。这项研究的结果证明了多种IRF-5剪接异构体的存在,它们在病毒介导的I型IFN基因诱导中具有不同的细胞类型特异性表达、细胞定位、差异调控和不同功能。