Kaposi's sarcoma-associated herpesviral IL-6 and human IL-6 open reading frames contain miRNA binding sites and are subject to cellular miRNA regulation
Kaposi's sarcoma‐associated herpesvirus (KSHV) encodes a viral interleukin 6 (vIL‐6) that mimics many activities of human IL‐6 (hIL‐6). Both vIL‐6 and hIL‐6 play important roles in stimulating the proliferation of tumours caused by KSHV. Here, we provide evidence that a miRNA pathway is involve...
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| Published in | The Journal of pathology Vol. 225; no. 3; pp. 378 - 389 |
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| Main Authors | , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Chichester, UK
John Wiley & Sons, Ltd
01.11.2011
Wiley |
| Subjects | |
| Online Access | Get full text |
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| Summary: | Kaposi's sarcoma‐associated herpesvirus (KSHV) encodes a viral interleukin 6 (vIL‐6) that mimics many activities of human IL‐6 (hIL‐6). Both vIL‐6 and hIL‐6 play important roles in stimulating the proliferation of tumours caused by KSHV. Here, we provide evidence that a miRNA pathway is involved in regulation of vIL‐6 and hIL‐6 expression through binding sites in their open reading frames (ORFs). We show a direct repression of vIL‐6 by hsa‐miR‐1293 and hIL‐6 by hsa‐miR‐608. The repression of vIL‐6 by miR‐1293 was reversed by disruption of the vIL‐6 miR‐1293 seed match through the introduction of point mutations. In addition, expression of vIL‐6 or hIL‐6 in KSHV‐infected cells could be enhanced by transfection of the respective miRNA inhibitors. In situ hybridization of human lymph node sections revealed that miR‐1293 is primarily expressed in the germinal centre but is deficient in the mantle zone of lymph nodes, where the expression of vIL‐6 is often found in patients with KSHV‐associated multicentric Castleman's disease, providing evidence of an anatomical correlation. Taking these factors together, our study indicates that IL‐6 expression can be regulated by miRNA interactions in its ORF and provides evidence for the role of these interactions in the pathogenesis of KSHV‐associated diseases. Copyright © 2011 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd. |
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| Bibliography: | Intramural Research Program of the National Institutes of Health Centre for Cancer Research istex:000C59D94285E5F234C07F6F4A53E541DB96790E ark:/67375/WNG-SKSMNFDF-G National Cancer Institute Supporting Information: Figure S1. Construction of vIL-6 and hIL-6 expression vectors. vIL-6 ORF (nt 17875-17264 of the KSHV genome, GenBank accession number U75698) and hIL-6 ORF (GenBank accession number NM_000600) were separately inserted into a p3xFLAG-CMV-14 vector (Sigma) at NotI and BamHI sites.Supporting Information: Figure S2. Copy numbers of miR-608 and miR-1293 in BCBL-1 cells with (+DOX) or without (−DOX) KSHV lytic induction. Doxycycline (DOX)-inducible TREx BCBL-1 RTA and vector control cell lines were used in the study. TREx BCBL1-RTA cells carrying an episomal KSHV genome and a tetracycline-inducible RTA (ORF50) expression vector were cultivated in the presence of doxycycline (1 µg/ml) to induce KSHV lytic infection. Copy numbers of miR-608 and miR-1293 were quantified by qRT-PCR on total small RNA extracted from the cells 24 h after DOX induction.Supporting Information: Figure S3. Distribution of endogenous miR-1293 and miR-155 in MCD lymph nodes. Panels show germinal center (GC) distribution of miR-1293 and mantle zone (MZ) distribution of miR-155. No miRNA, no specific miRNA probe. See more details for in situ hybridization in Figure 7. Scale bar represents 200 Φm.Supporting Information ArticleID:PATH2962 Conflict of interest statement. Dr Giovanna Tosato, the spouse of one of the authors, RY, is a co-inventor on a patent describing the measurement of KSHV vIL-6. This invention was made when GT was an employee of the US Government under Code of Federal Regulations 45, Part 7. All rights, title and interest to this patent have been assigned to the US Department of Health and Human Services. The government conveys a portion of the royalties it receives to its employee inventors under the Federal Technology Transfer Act of 1986 (PL 99-502). Conflict of interest statement. Dr Giovanna Tosato, the spouse of one of the authors, RY, is a co‐inventor on a patent describing the measurement of KSHV vIL‐6. This invention was made when GT was an employee of the US Government under Code of Federal Regulations 45, Part 7. All rights, title and interest to this patent have been assigned to the US Department of Health and Human Services. The government conveys a portion of the royalties it receives to its employee inventors under the Federal Technology Transfer Act of 1986 (PL 99–502). ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2 |
| ISSN: | 0022-3417 1096-9896 1096-9896 |
| DOI: | 10.1002/path.2962 |