Histone H2B-IFI16 Recognition of Nuclear Herpesviral Genome Induces Cytoplasmic Interferon-β Responses.
Histone H2B-IFI16 Recognition of Nuclear Herpesviral Genome Induces Cytoplasmic Interferon-β Responses.
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组蛋白 H2B-IFI16 识别核疱疹病毒基因组诱导细胞质干扰素-β反应
DOI：
10.1371/journal.ppat.1005967
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发表时间：
2016-10
期刊：
影响因子：
6.7
通讯作者：
Chandran B
中科院分区：
文献类型：
--
作者：
Iqbal J;Ansari MA;Kumar B;Dutta D;Roy A;Chikoti L;Pisano G;Dutta S;Vahedi S;Veettil MV;Chandran B
IFI 16（γ-干扰素诱导蛋白16）是一种主要参与转录调控的核蛋白，也作为先天免疫应答DNA传感器发挥作用，并诱导IL-1β和抗病毒1型干扰素-β（IFN-β）细胞因子。我们已经表明，IFI 16，与BRCA 1，作为一个序列独立的核传感器的附加型双链DNA基因组的KSHV，EBV和HSV-1。这些疱疹病毒基因组的识别导致IFI 16乙酰化、BRCA 1-IFI 16-ASC-procaspase-1炎性小体形成、细胞质易位和IL-1β产生。乙酰化的IFI 16还与细胞质STING相互作用并诱导IFN-β。然而，参与STING激活的IFI 16相关核蛋白的身份及其机制尚不清楚。从未感染的内皮细胞核裂解物的抗IFI 16抗体沉淀的蛋白质的质谱显示，组蛋白H2 B与IFI 16相互作用。单和双邻近连接显微镜，免疫沉淀，EdU-基因组标记的病毒感染，和染色质免疫沉淀研究表明，H2 B与IFI 16和BRCA 1在细胞核中的生理条件。新生KSHV和HSV-1感染以及潜伏KSHV和EBV感染诱导H2 B-IFI 16、H2 B-BRCA 1和IFI 16-ASC复合物的细胞质分布。痘苗病毒（dsDNA）的胞质复制不诱导核内H2 B-IFI 16或H2 B向胞质内的再分布。H2 B在KSHV和HSV-1基因组识别过程中起关键作用。通过IFI 16-H2 B-BRCA 1的病毒基因组感应导致BRCA 1依赖性的p300募集以及H2 B和IFI 16的乙酰化。BRCA 1敲除或p300抑制废除了H2 B-IFI 16或H2 B的乙酰化。Ran-GTP蛋白介导乙酰化H2 B和IFI 16与BRCA 1一起沿着移位至细胞质，而不依赖于IFI 16-ASC炎性体。ASC基因敲除不影响H2 B的乙酰化、胞质转运以及STING与IFI 16和H2 B的结合。H2 B的缺乏不影响IFI 16-ASC结合和胞质分布，因此证明IFI 16-H2 B复合物不依赖于感染期间形成的IFI 16-ASC-半胱氨酸天冬氨酸蛋白酶原-1-炎性体复合物。H2 B-IFI 16-BRCA 1复合物与细胞质中的cGAS和STING相互作用，导致TBK 1和IRF 3磷酸化、pIRF 3的核转位和IFN-β产生。H2 B、cGAS和STING的沉默抑制IFN-β诱导但不抑制IL-1β分泌，并且在感染期间通过H2 B和IFI 16敲低显著降低cGAMP活性。在KSHV和HSV-1从头感染期间，ASC的沉默抑制IL-1β分泌，但不抑制IFN-β分泌。这些研究将H2 B鉴定为介导新的染色体外功能的先天核传感器，并揭示了两种IFI 16复合物介导KSHV和HSV-1基因组识别应答，其中IFI 16-BRCA 1-H2 B复合物的识别导致IFN-β应答，IFI 16-BRCA 1的识别导致炎性小体应答。真核细胞引发针对包括病毒在内的入侵微生物的先天免疫应答。IFI 16是一种主要的核蛋白，已成为一种先天性反应核DNA传感器。IFI 16-BRCA 1识别核KSHV、HSV-1和EBV dsDNA基因组导致IFI 16乙酰化、BRCA 1-IFI 16-ASC-半胱氨酸天冬氨酸蛋白酶原-1炎性体复合物的细胞质易位和IL-1β产生。在这里，我们证明，组蛋白H2 B与IFI 16-BRCA 1在细胞核中的生理条件下。IFI 16-H2 B-BRCA 1识别核病毒基因组导致BRCA 1-p300介导的H2 B和IFI 16的乙酰化，以及H2 B-IFI 16-BRCA 1通过Ran GTP蛋白质的胞质转运。炎性体非依赖性细胞质IFI 16-H2 B-BRCA 1复合物与cGAS和STING相互作用，导致TBK 1和IRF 3磷酸化，以及核pIRF 3介导的IFN-β诱导。H2 B敲低抑制IFN-β产生，而ASC沉默不影响IFN-β诱导。我们的研究将H2 B鉴定为先天性核传感器，并揭示两种IFI 16复合物介导核疱疹病毒基因组识别反应，IFI 16-BRCA 1-H2 B-IFN-β反应和IFI 16-BRCA 1-炎性体反应。
IFI16 (gamma-interferon-inducible protein 16), a predominantly nuclear protein involved in transcriptional regulation, also functions as an innate immune response DNA sensor and induces the IL-1β and antiviral type-1 interferon-β (IFN-β) cytokines. We have shown that IFI16, in association with BRCA1, functions as a sequence independent nuclear sensor of episomal dsDNA genomes of KSHV, EBV and HSV-1. Recognition of these herpesvirus genomes resulted in IFI16 acetylation, BRCA1-IFI16-ASC-procaspase-1 inflammasome formation, cytoplasmic translocation, and IL-1β generation. Acetylated IFI16 also interacted with cytoplasmic STING and induced IFN-β. However, the identity of IFI16 associated nuclear proteins involved in STING activation and the mechanism is not known. Mass spectrometry of proteins precipitated by anti-IFI16 antibodies from uninfected endothelial cell nuclear lysate revealed that histone H2B interacts with IFI16. Single and double proximity ligation microscopy, immunoprecipitation, EdU-genome labeled virus infection, and chromatin immunoprecipitation studies demonstrated that H2B is associated with IFI16 and BRCA1 in the nucleus in physiological conditions. De novo KSHV and HSV-1 infection as well as latent KSHV and EBV infection induces the cytoplasmic distribution of H2B-IFI16, H2B-BRCA1 and IFI16-ASC complexes. Vaccinia virus (dsDNA) cytoplasmic replication didn’t induce the redistribution of nuclear H2B-IFI16 or H2B into the cytoplasm. H2B is critical in KSHV and HSV-1 genome recognition by IFI16 during de novo infection. Viral genome sensing by IFI16-H2B-BRCA1 leads to BRCA1 dependent recruitment of p300, and acetylation of H2B and IFI16. BRCA1 knockdown or inhibition of p300 abrogated the acetylation of H2B-IFI16 or H2B. Ran-GTP protein mediated the translocation of acetylated H2B and IFI16 to the cytoplasm along with BRCA1 that is independent of IFI16-ASC inflammasome. ASC knockdown didn’t affect the acetylation of H2B, its cytoplasmic transportation, and the association of STING with IFI16 and H2B during KSHV infection. Absence of H2B didn’t affect IFI16-ASC association and cytoplasmic distribution and thus demonstrating that IFI16-H2B complex is independent of IFI16-ASC-procaspase-1-inflammasome complex formed during infection. The H2B-IFI16-BRCA1 complex interacted with cGAS and STING in the cytoplasm leading to TBK1 and IRF3 phosphorylation, nuclear translocation of pIRF3 and IFN-β production. Silencing of H2B, cGAS and STING inhibited IFN-β induction but not IL-1β secretion, and cGAMP activity is significantly reduced by H2B and IFI16 knockdown during infection. Silencing of ASC inhibited IL-1β secretion but not IFN-β secretion during de novo KSHV and HSV-1 infection. These studies identify H2B as an innate nuclear sensor mediating a novel extra chromosomal function, and reveal that two IFI16 complexes mediate KSHV and HSV-1 genome recognition responses, with recognition by the IFI16-BRCA1-H2B complex resulting in IFN-β responses and recognition by IFI16-BRCA1 resulting in inflammasome responses. Eukaryotic cells elicit innate immune responses against invading microbes including viruses. IFI16, a predominantly nuclear protein, has emerged as an innate response nuclear DNA sensor. Recognition of nuclear KSHV, HSV-1 and EBV dsDNA genomes by IFI16-BRCA1 leads to IFI16 acetylation, cytoplasmic translocation of the BRCA1-IFI16-ASC-procaspase-1 inflammasome complex and IL-1β generation. Here, we demonstrate that histone H2B is associated with IFI16-BRCA1 in the nucleus under physiological conditions. Recognition of nuclear viral genomes by IFI16-H2B-BRCA1 leads to BRCA1-p300 mediated acetylation of H2B and IFI16, and cytoplasmic transport of H2B-IFI16-BRCA1 via Ran GTP protein. The inflammasome independent cytoplasmic IFI16-H2B-BRCA1 complex interacts with cGAS and STING resulting in TBK1 and IRF3 phosphorylation, and nuclear pIRF3-mediated IFN-β induction. H2B knockdown inhibits IFN-β production while ASC silencing doesn’t affect IFN-β induction. Our studies identify H2B as an innate nuclear sensor and reveal that two IFI16 complexes mediate nuclear herpesviral genome recognition responses, IFI16-BRCA1-H2B-IFN-β responses and IFI16-BRCA1-inflammasome responses.
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影响因子：
6.7
作者：
Ansari MA;Dutta S;Veettil MV;Dutta D;Iqbal J;Kumar B;Roy A;Chikoti L;Singh VV;Chandran B
通讯作者：
Chandran B
影响因子：
3.7
作者：
Kobiyama K;Kawashima A;Jounai N;Takeshita F;Ishii KJ;Ito T;Suzuki K
通讯作者：
Suzuki K
影响因子：
6.7
作者：
Johnson KE;Bottero V;Flaherty S;Dutta S;Singh VV;Chandran B
通讯作者：
Chandran B
DOI：
10.1083/jcb.153.2.251
发表时间：
2001-04-16
期刊：
The Journal of cell biology
影响因子：
--
作者：
Mosammaparast N;Jackson KR;Guo Y;Brame CJ;Shabanowitz J;Hunt DF;Pemberton LF
通讯作者：
Pemberton LF
影响因子：
30.3
作者：
Kerur N;Veettil MV;Sharma-Walia N;Bottero V;Sadagopan S;Otageri P;Chandran B
通讯作者：
Chandran B