TY - JOUR
T1 - Epstein-Barr virus microRNAs regulate B cell receptor signal transduction and lytic reactivation
AU - Chen, Yan
AU - Fachko, Devin
AU - Ivanov, Nikita S.
AU - Skinner, Camille M.
AU - Skalsky, Rebecca L.
N1 - Funding Information:
This work was supported by a Pathway to Independence Award CA175181 from the National Cancer Institute and a pilot award from the Collins Medical Trust to RLS. CMS was additionally supported by funds from the Community Engagement and Leadership in Science (CELS) Program at Lewis and Clark College. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors thank Isabella Brink and Robert Manni, both previous undergraduate students in the Skalsky lab, for assistance with molecular cloning, and Dr. Lindsey Crawford at VGTI for assistance with cell sorting.
Publisher Copyright:
© 2019 Chen et al.
PY - 2019
Y1 - 2019
N2 - MicroRNAs (miRNAs) are post-transcriptional regulatory RNAs that can modulate cell signaling and play key roles in cell state transitions. Epstein-Barr virus (EBV) expresses >40 viral miRNAs that manipulate both viral and cellular gene expression patterns and contribute to reprogramming of the host environment during infection. Here, we identified a subset of EBV miRNAs that desensitize cells to B cell receptor (BCR) stimuli, and attenuate the downstream activation of NF-kappaB or AP1-dependent transcription. Bioinformatics and pathway analysis of Ago PAR-CLIP datasets identified multiple EBV miRNA targets related to BCR signal transduction, including GRB2, SOS1, MALT1, RAC1, and INPP5D, which we validated in reporter assays. BCR signaling is critical for B cell activation, proliferation, and differentiation, and for EBV, is linked to reactivation. In functional assays, we demonstrate that EBV miR-BHRF1-2-5p contributes to the growth of latently infected B cells through GRB2 regulation. We further determined that activities of EBV miR-BHRF1-2-5p, EBV miR-BART2-5p, and a cellular miRNA, miR-17-5p, directly regulate virus reactivation triggered by BCR engagement. Our findings provide mechanistic insight into some of the key miRNA interactions impacting the proliferation of latently infected B cells and importantly, governing the latent to lytic switch.
AB - MicroRNAs (miRNAs) are post-transcriptional regulatory RNAs that can modulate cell signaling and play key roles in cell state transitions. Epstein-Barr virus (EBV) expresses >40 viral miRNAs that manipulate both viral and cellular gene expression patterns and contribute to reprogramming of the host environment during infection. Here, we identified a subset of EBV miRNAs that desensitize cells to B cell receptor (BCR) stimuli, and attenuate the downstream activation of NF-kappaB or AP1-dependent transcription. Bioinformatics and pathway analysis of Ago PAR-CLIP datasets identified multiple EBV miRNA targets related to BCR signal transduction, including GRB2, SOS1, MALT1, RAC1, and INPP5D, which we validated in reporter assays. BCR signaling is critical for B cell activation, proliferation, and differentiation, and for EBV, is linked to reactivation. In functional assays, we demonstrate that EBV miR-BHRF1-2-5p contributes to the growth of latently infected B cells through GRB2 regulation. We further determined that activities of EBV miR-BHRF1-2-5p, EBV miR-BART2-5p, and a cellular miRNA, miR-17-5p, directly regulate virus reactivation triggered by BCR engagement. Our findings provide mechanistic insight into some of the key miRNA interactions impacting the proliferation of latently infected B cells and importantly, governing the latent to lytic switch.
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U2 - 10.1371/journal.ppat.1007535
DO - 10.1371/journal.ppat.1007535
M3 - Article
C2 - 30615681
AN - SCOPUS:85060140072
SN - 1553-7366
VL - 15
JO - PLoS pathogens
JF - PLoS pathogens
IS - 1
M1 - e1007535
ER -