Systematic Analysis of Viral and Cellular MicroRNA Targets in Cells Latently Infected with Human γ-Herpesviruses by RISC Immunoprecipitation Assay

被引：188

作者：

Doelken, Lars ^{[1
]}

Malterer, Georg ^{[1
]}

Erhard, Florian ^{[2
]}

Kothe, Sheila ^{[1
]}

Friedel, Caroline C. ^{[2
]}

Suffert, Guillaume ^{[3
]}

Marcinowski, Lisa ^{[1
]}

Motsch, Natalie ^{[4
]}

Barth, Stephanie ^{[4
]}

Beitzinger, Michaela ^{[5
]}

Lieber, Diana ^{[1
]}

Bailer, Susanne M. ^{[1
]}

Hoffmann, Reinhard ^{[6
]}

Ruzsics, Zsolt ^{[1
]}

Kremmer, Elisabeth ^{[7
]}

Pfeffer, Sebastien ^{[3
]}

Zimmer, Ralf ^{[2
]}

Koszinowski, Ulrich H. ^{[1
]}

Grasser, Friedrich ^{[4
]}

Meister, Gunter ^{[5
]}

Haas, Juergen ^{[1
,8
]}

机构：

[1] Univ Munich, Max von Pettenkofer Inst, D-80336 Munich, Germany

[2] Univ Munich, Inst Informat, D-80333 Munich, Germany

[3] Univ Strasbourg, CNRS, Inst Biol Mol & Cellulaire, F-67084 Strasbourg, France

[4] Univ Klinikum Saarlandes, Inst Virol, D-66421 Homburg, Germany

[5] Max Planck Inst Biochem, D-82152 Martinsried, Germany

[6] Tech Univ Munich, Inst Med Microbiol, D-81675 Munich, Germany

[7] Inst Mol Immunol, Helmholtz Zentrum Munchen, D-81377 Munich, Germany

[8] Univ Edinburgh, Div Pathway Med, Edinburgh EH16 4SB, Midlothian, Scotland

来源：

CELL HOST & MICROBE | 2010年 / 7卷 / 04期

关键词：

SARCOMA-ASSOCIATED HERPESVIRUS; PRIMARY EFFUSION LYMPHOMA; VIRUS-ENCODED MICRORNAS; OUTER-MEMBRANE; MESSENGER-RNAS; RIP-CHIP; B-CELLS; IDENTIFICATION; EXPRESSION; PATHWAY;

D O I：

10.1016/j.chom.2010.03.008

中图分类号：

Q93 [微生物学];

学科分类号：

071005 [微生物学];

摘要：

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 sarcoma-associated 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 IPO7) 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.

引用

页码：324 / 334

页数：11

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