TY - JOUR
T1 - Systematic analysis of viral and cellular microRNA targets in cells latently infected with human γ-herpesviruses by RISC immunoprecipitation assay
AU - Dölken, Lars
AU - Malterer, Georg
AU - Erhard, Florian
AU - Kothe, Sheila
AU - Friedel, Caroline C.
AU - Suffert, Guillaume
AU - Marcinowski, Lisa
AU - Motsch, Natalie
AU - Barth, Stephanie
AU - Beitzinger, Michaela
AU - Lieber, Diana
AU - Bailer, Susanne M.
AU - Hoffmann, Reinhard
AU - Ruzsics, Zsolt
AU - Kremmer, Elisabeth
AU - Pfeffer, Sébastien
AU - Zimmer, Ralf
AU - Koszinowski, Ulrich H.
AU - Grässer, Friedrich
AU - Meister, Gunter
AU - Haas, Jurgen
PY - 2010/4/22
Y1 - 2010/4/22
N2 - The mRNA targets of microRNAs (miRNAs) can be identified by immunoprecipitation of Argonaute (Ago) protein-containing RNA-induced silencing complexes (RISCs) followed by microarray analysis (RIP-Chip). Here we used Ago2-based RIP-Chip to identify transcripts targeted by Kaposi's sarcomaassociated herpesvirus (KSHV) miRNAs (n = 114), Epstein-Barr virus (EBV) miRNAs (n = 44), and cellular miRNAs (n = 2337) in six latently infected or stably transduced human B cell lines. Of the six KSHV miRNA targets chosen for validation, four showed regulation via their 3'UTR, while two showed regulation via binding sites within coding sequences. Two genes governing cellular transport processes (TOMM22 and IP07) were confirmed to be targeted by EBV miRNAs. A significant number of viral miRNA targets were upregulated in infected cells, suggesting that viral miRNAs preferentially target cellular genes induced upon infection. Transcript half-life both of cellular and viral miRNA targets negatively correlated with recruitment to RISC complexes, indicating that RIP-Chip offers a quantitative estimate of miRNA function.
AB - The mRNA targets of microRNAs (miRNAs) can be identified by immunoprecipitation of Argonaute (Ago) protein-containing RNA-induced silencing complexes (RISCs) followed by microarray analysis (RIP-Chip). Here we used Ago2-based RIP-Chip to identify transcripts targeted by Kaposi's sarcomaassociated herpesvirus (KSHV) miRNAs (n = 114), Epstein-Barr virus (EBV) miRNAs (n = 44), and cellular miRNAs (n = 2337) in six latently infected or stably transduced human B cell lines. Of the six KSHV miRNA targets chosen for validation, four showed regulation via their 3'UTR, while two showed regulation via binding sites within coding sequences. Two genes governing cellular transport processes (TOMM22 and IP07) were confirmed to be targeted by EBV miRNAs. A significant number of viral miRNA targets were upregulated in infected cells, suggesting that viral miRNAs preferentially target cellular genes induced upon infection. Transcript half-life both of cellular and viral miRNA targets negatively correlated with recruitment to RISC complexes, indicating that RIP-Chip offers a quantitative estimate of miRNA function.
UR - https://www.scopus.com/pages/publications/77954573536
U2 - 10.1016/j.chom.2010.03.008
DO - 10.1016/j.chom.2010.03.008
M3 - Article
C2 - 20413099
AN - SCOPUS:77954573536
SN - 1931-3128
VL - 7
SP - 324
EP - 334
JO - Cell Host and Microbe
JF - Cell Host and Microbe
IS - 4
ER -