DNA-PK/Ku complex binds to latency-associated nuclear antigen and negatively regulates Kaposi’s sarcoma-associated herpesvirus latent replication

Cha, Seho; Lim, Chunghun; Lee, Jae Young; Song, Young-Bo; Park, Junsoo; Choe, Joonho; Seo, Taegun

doi:10.1016/j.bbrc.2010.03.086

Cited by 19 publications

(26 citation statements)

References 41 publications

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“…In our TAP-MS screening, a few novel proteins that were related to transcriptional regulation and posttranslational modification were identified, such as KAP1, NCOR2, retinoblastoma-associated protein (RBBP), SMAD, USP7, and SUMO3. Previously reported LANA-interacting proteins, like BRD2 (54), SIN3A (30), PARP1 (67), PRKDC (DNA-PK) (68), XRCC6 (KU70) (68), and KDM3A (48), were also found in this study (Table 2). We validated the interaction between LANA and KAP1 in vivo and in vitro.…”

Section: Discussionsupporting

confidence: 82%

“…In this study, we showed that LANA interacted with the KAP1 protein and repressed lytic gene expression to facilitate the establishment of KSHV latency. Multiple proteins have been shown to interact with LANA (26,30,38,45,(48)(49)(50)(53)(54)(55)(56)(57)(58)(59)(60)(61)(62)(63)(64)(65)(66)(67)(68)(69). In our TAP-MS screening, a few novel proteins that were related to transcriptional regulation and posttranslational modification were identified, such as KAP1, NCOR2, retinoblastoma-associated protein (RBBP), SMAD, USP7, and SUMO3.…”

Section: Discussionmentioning

confidence: 99%

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Kaposi's Sarcoma-Associated Herpesvirus-Encoded LANA Interacts with Host KAP1 To Facilitate Establishment of Viral Latency

Sun

Liang

Gao

et al. 2014

J Virol

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Kaposi's sarcoma-associated herpesvirus (KSHV) typically displays two different phases in its life cycle, the default latent phase and the lytic phase. There is a short period of lytic gene expression in the early stage of KSHV primary infection. The factors involved in the shutdown process of lytic gene expression are poorly identified. It has been shown that the latency-associated nuclear antigen (LANA) encoded by KSHV plays an important role in the establishment of viral latency. In screening, we identified a host protein, Krüppel-associated box domain-associated protein 1 (KAP1), that bound to LANA. We validated the interaction between LANA and KAP1 in vivo and in vitro, as well as their colocalization in the nucleus. We mapped out that LANA interacted with both the N-and C-terminal domains of KAP1. Based on the interface of LANA-KAP1 interaction determined, we proved that LANA recruited KAP1 to the RTA promoter region of the KSHV genome. We revealed that KAP1 was involved in transcriptional repression by LANA. We found multiple cooccupation sites of LANA and KAP1 on the whole KSHV genome by chromatin immunoprecipitation for sequencing (ChIP-seq) and demonstrated that LANA-recruited KAP1 played a critical role in the shutdown of lytic gene expression during the early stage of KSHV primary infection. Taken together, our data suggest that LANA interacts with KAP1 and represses lytic gene expression to facilitate the establishment of KSHV latency. IMPORTANCEOur study revealed the mechanism of transcriptional repression by LANA during KSHV primary infection, providing new insights into the process of KSHV latency establishment.

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Section: Discussionsupporting

confidence: 82%

Section: Discussionmentioning

confidence: 99%

Kaposi's Sarcoma-Associated Herpesvirus-Encoded LANA Interacts with Host KAP1 To Facilitate Establishment of Viral Latency

Sun

Liang

Gao

et al. 2014

J Virol

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“…Coupled with the ability of the RFHVMn and KSHV LANA N-terminal domains to interact directly with importin β1, this suggests an underlying ability of the RV1 LANA homologs to interact with nucleolar components. Interestingly, the 52 N-terminal amino acids of KSHV LANA bound to nucleolar proteins hnRNP A1, B23.1, the DNA-dependent protein kinase (DNA-PK) subunits Ku86 and Ku70, and YB-1 in GST-pulldown experiments (Cha et al, 2010). …”

Section: Discussionmentioning

confidence: 99%

ORF73 LANA homologs of RRV and MneRV2 contain an extended RGG/RG-rich nuclear and nucleolar localization signal that interacts directly with importin β1 for non-classical nuclear import

2017

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The latency-associated nuclear antigens (LANA) of KSHV and macaque RFHVMn, members of the RV1 rhadinovirus lineage, are closely related with conservation of complex nuclear localization signals (NLS) containing bipartite KR-rich motifs and RG-rich domains, which interact distinctly with importins α and β1 for nuclear import via classical and non-classical pathways, respectively. RV1 LANAs are expressed in the nucleus of latently-infected cells where they inhibit replication and establish a dominant RV1 latency. Here we show that LANA homologs of macaque RRV and MneRV2 from the more distantly-related RV2 lineage, lack the KR-rich NLS, and instead have a large RG-rich NLS with multiple RG dipeptides and a conserved RGG motif. The RG-NLS interacts uniquely with importin β1, which mediates nuclear import and accumulation of RV2 LANA in the nucleolus. The alternative nuclear import and localization of RV2 LANA homologs may contribute to the dominant RV2 lytic replication phenotype.

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“…Ser10, which is within the chromatin binding domain, was previously shown to be phosphorylated by casein kinase 1 (97). Phosphorylation of the N-terminal (aa 91 to 340) and C-terminal (aa 906 to 1070; BCBL-1 LANA sequence) regions of LANA has been previously detected (98,99), and our mass spectrometry analysis identified the phosphorylation of serines 186, 187, 1027, 1041, 1051, and 1067 and threonines 960 and 1054 in these regions (see Fig. S2 in the supplemental material).…”

Section: Discussionmentioning

confidence: 99%

Full-Length Isoforms of Kaposi's Sarcoma-Associated Herpesvirus Latency-Associated Nuclear Antigen Accumulate in the Cytoplasm of Cells Undergoing the Lytic Cycle of Replication

Garrigues

Howard

Barcy

et al. 2017

J Virol

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The latency-associated nuclear antigen (LANA) of the Kaposi's sarcomaassociated herpesvirus (KSHV) performs a variety of functions to establish and maintain KSHV latency. During latency, LANA localizes to discrete punctate spots in the nucleus, where it tethers viral episomes to cellular chromatin and interacts with nuclear components to regulate cellular and viral gene expression. Using highly sensitive tyramide signal amplification, we determined that LANA localizes to the cytoplasm in different cell types undergoing the lytic cycle of replication after de novo primary infection and after spontaneous, tetradecanoyl phorbol acetate-, or open reading frame 50 (ORF50)/replication transactivator (RTA)-induced activation. We confirmed the presence of cytoplasmic LANA in a subset of cells in lytically active multicentric Castleman disease lesions. The induction of cellular migration by scratch-wounding confluent cell cultures, culturing under subconfluent conditions, or induction of cell differentiation in primary cultures upregulated the number of cells permissive for primary lytic KSHV infection. The induction of lytic replication was characterized by high-level expression of cytoplasmic LANA and nuclear ORF59, a marker of lytic replication. Subcellular fractionation studies revealed the presence of multiple isoforms of LANA in the cytoplasm of ORF50/RTA-activated Vero cells undergoing primary infection. Mass spectrometry analysis demonstrated that cytoplasmic LANA isoforms were full length, containing the N-terminal nuclear localization signal. These results suggest that trafficking of LANA to different subcellular locations is a regulated phenomenon, which allows LANA to interact with cellular components in different compartments during both the latent and the replicative stages of the KSHV life cycle. IMPORTANCE Kaposi's sarcoma-associated herpesvirus (KSHV) causes AIDS-related malignancies, including lymphomas and Kaposi's sarcoma. KSHV establishes lifelong infections using its latency-associated nuclear antigen (LANA). During latency, LANA localizes to the nucleus, where it connects viral and cellular DNA complexes and regulates gene expression, allowing the virus to maintain long-term infections. Our research shows that intact LANA traffics to the cytoplasm of cells undergoing permissive lytic infections and latently infected cells in which the virus is induced to replicate. This suggests that LANA plays important roles in the cytoplasm and nuclear compartments of the cell during different stages of the KSHV life cycle. Determining cytoplasmic function and mechanism for regulation of the nuclear localiza-

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DNA-PK/Ku complex binds to latency-associated nuclear antigen and negatively regulates Kaposi’s sarcoma-associated herpesvirus latent replication

Cited by 19 publications

References 41 publications

Kaposi's Sarcoma-Associated Herpesvirus-Encoded LANA Interacts with Host KAP1 To Facilitate Establishment of Viral Latency

Kaposi's Sarcoma-Associated Herpesvirus-Encoded LANA Interacts with Host KAP1 To Facilitate Establishment of Viral Latency

ORF73 LANA homologs of RRV and MneRV2 contain an extended RGG/RG-rich nuclear and nucleolar localization signal that interacts directly with importin β1 for non-classical nuclear import

Full-Length Isoforms of Kaposi's Sarcoma-Associated Herpesvirus Latency-Associated Nuclear Antigen Accumulate in the Cytoplasm of Cells Undergoing the Lytic Cycle of Replication

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