Kaposi's Sarcoma Herpesvirus microRNAs Target Caspase 3 and Regulate Apoptosis

PLoS Pathogens

Maud Contrant¹  Aurélie Fender¹  Guillaume Suffert¹  Mihaela Zavolan²  Jean Hausser²  Juergen G. Haas³  Vladimir Benes⁴  Tomi Ivacevic⁴  Johanna Viiliäinen⁵  Päivi M. Ojala⁵  Frédéric Gros⁶  Olivier Voinnet⁷  Georg Malterer⁸  Sébastien Pfeffer⁸ 

[1] Architecture et Réactivité de l'ARN, Institut de Biologie Moléculaire et Cellulaire du CNRS, Université de Strasbourg, Strasbourg, France;Biozentrum der Universität Basel and Swiss Institute of Bioinformatics, Basel, Switzerland;Foundation for the Finnish Cancer Institute, Helsinki, Finland;GeneCore (Genomics Core Facility), EMBL, Heidelberg, Germany;Genome-Scale Biology Program, Biomedicum Helsinki and Institute of Biomedicine, University of Helsinki, Helsinki, Finland;Immunologie et Chimie Thérapeutiques UPR 9021, Institut de Biologie Moléculaire et Cellulaire du CNRS, Université de Strasbourg, Strasbourg, France;Institut de Biologie Moléculaire des Plantes du CNRS, Strasbourg, France;Max von Pettenkofer-Institute, Ludwig-Maximilians-University Munich, Munich, Germany

关键词: MicroRNAs;    Apoptosis;    Kaposi's sarcoma-associated herpesvirus;    Untranslated regions;    Gene expression;    Luciferase assay;    Luciferase;    Microarrays;   

DOI  :  10.1371/journal.ppat.1002405

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Kaposi's sarcoma herpesvirus (KSHV) encodes a cluster of twelve micro (mi)RNAs, which are abundantly expressed during both latent and lytic infection. Previous studies reported that KSHV is able to inhibit apoptosis during latent infection; we thus tested the involvement of viral miRNAs in this process. We found that both HEK293 epithelial cells and DG75 cells stably expressing KSHV miRNAs were protected from apoptosis. Potential cellular targets that were significantly down-regulated upon KSHV miRNAs expression were identified by microarray profiling. Among them, we validated by luciferase reporter assays, quantitative PCR and western blotting caspase 3 (Casp3), a critical factor for the control of apoptosis. Using site-directed mutagenesis, we found that three KSHV miRNAs, miR-K12-1, 3 and 4-3p, were responsible for the targeting of Casp3. Specific inhibition of these miRNAs in KSHV-infected cells resulted in increased expression levels of endogenous Casp3 and enhanced apoptosis. Altogether, our results suggest that KSHV miRNAs directly participate in the previously reported inhibition of apoptosis by the virus, and are thus likely to play a role in KSHV-induced oncogenesis.

【 授权许可】

CC BY