@article{eb54127af1124b0b9eb1f03326d0e162,
title = "KRAS Engages AGO2 to Enhance Cellular Transformation",
abstract = "Oncogenic mutations in RAS provide a compelling yet intractable therapeutic target. Using co-immunoprecipitation mass spectrometry, we uncovered an interaction between RAS and Argonaute 2 (AGO2). Endogenously, RAS and AGO2 co-sediment and co-localize in the endoplasmic reticulum. The AGO2 N-terminal domain directly binds the Switch II region of KRAS, agnostic of nucleotide (GDP/GTP) binding. Functionally, AGO2 knockdown attenuates cell proliferation in mutant KRAS-dependent cells and AGO2 overexpression enhances KRASG12V-mediated transformation. Using AGO2-/- cells, we demonstrate that the RAS-AGO2 interaction is required for maximal mutant KRAS expression and cellular transformation. Mechanistically, oncogenic KRAS attenuates AGO2-mediated gene silencing. Overall, the functional interaction with AGO2 extends KRAS function beyond its canonical role in signaling.",
keywords = "Argonaute 2, Cancer, EIF2C2, KRAS, RNA silencing",
author = "Sunita Shankar and Sethuramasundaram Pitchiaya and Rohit Malik and Vishal Kothari and Yasuyuki Hosono and Yocum, {Anastasia K.} and Harika Gundlapalli and Yasmine White and Ari Firestone and Xuhong Cao and Dhanasekaran, {Saravana M.} and Stuckey, {Jeanne A.} and Gideon Bollag and Kevin Shannon and Walter, {Nils G.} and Chandan Kumar-Sinha and Chinnaiyan, {Arul M.}",
note = "Funding Information: We acknowledge the work of Shanker Kalyan-Sundaram, Krishnapriya Chinnaswamy, Vijaya L. Dommeti, Matthew Shuler, Anton Poliakov, Xiaoju Wang, and Vishalakshi Krishnan, who helped with analysis and experimentation. We thank Phillip Zamore for providing AGO2−/− and AGO2−/− + AGO2 MEFs and Mariano Barbacid for providing KRAS-only-expressing MEFs. We thank Eric Fearon for helpful discussions; Joseph Mierzwa, Kevin Eid, and Jincheng Pan for technical assistance; Bushra Ateeq and Rachell Stender for help with the xenograft studies; William Brown for His-AGO2 protein preparation; and Robin Kunkel for assistance with schematic representations. We also thank Ingrid Apel, Xiaojun Jing, and David O. Apiyo (Pall Life Sciences) for carrying out additional experiments that were not used for the final manuscript. We also benefited from discussions with Denzil Bernard (structure-function) and John O{\textquoteright}Bryan (University of Illinois; nucleotide loading). We thank Ester Fernandez-Salas for her inputs on the manuscript. We thank the University of Michigan Xenograft Core and Dr. Diane Simeone for providing PDX1319 cell line. S.P. was supported by IFOM Fondazione Istituto FIRC di Oncologia Molecolare, Milan, Italy (sponsor: Fabrizio D{\textquoteright}Adda di Fagagna). R.M. is supported by Prostate Cancer Foundation Young Investigator Award. Y.W. was supported by a Howard Hughes Medical Institute (HHMI) Medical Student Research Fellowship. and A.F. by a Damon Runyon Foundation Fellowship. A.M.C. is supported by the Alfred A. Taubman Institute and the HHMI. A.M.C. and K.S. are American Cancer Society Research Professors. This project is supported in part by NIH grants NIH 1R21 AI109791 (PI: N.G.W.), RO1 CA154365 and R37 CA40046, and the Prostate Cancer Foundation (PI: A.M.C.). Funding Information: We acknowledge the work of Shanker Kalyan-Sundaram, Krishnapriya Chinnaswamy, Vijaya L. Dommeti, Matthew Shuler, Anton Poliakov, Xiaoju Wang, and Vishalakshi Krishnan, who helped with analysis and experimentation. We thank Phillip Zamore for providing AGO2-/- and AGO2-/- + AGO2 MEFs and Mariano Barbacid for providing KRAS-only-expressing MEFs. We thank Eric Fearon for helpful discussions; Joseph Mierzwa, Kevin Eid, and Jincheng Pan for technical assistance; Bushra Ateeq and Rachell Stender for help with the xenograft studies; William Brown for His-AGO2 protein preparation; and Robin Kunkel for assistance with schematic representations. We also thank Ingrid Apel, Xiaojun Jing, and David O. Apiyo (Pall Life Sciences) for carrying out additional experiments that were not used for the final manuscript. We also benefited from discussions with Denzil Bernard (structure-function) and John O{\textquoteright}Bryan (University of Illinois; nucleotide loading). We thank Ester Fernandez-Salas for her inputs on the manuscript. We thank the University of Michigan Xenograft Core and Dr. Diane Simeone for providing PDX1319 cell line. S.P. was supported by IFOM Fondazione Istituto FIRC di Oncologia Molecolare, Milan, Italy (sponsor: Fabrizio D{\textquoteright}Adda di Fagagna). R.M. is supported by Prostate Cancer Foundation Young Investigator Award. Y.W. was supported by a Howard Hughes Medical Institute (HHMI) Medical Student Research Fellowship. and A.F. by a Damon Runyon Foundation Fellowship. A.M.C. is supported by the Alfred A. Taubman Institute and the HHMI. A.M.C. and K.S. are American Cancer Society Research Professors. This project is supported in part by NIH grants NIH 1R21 AI109791 (PI: N.G.W.), RO1 CA154365 and R37 CA40046, and the Prostate Cancer Foundation (PI: A.M.C.). Publisher Copyright: {\textcopyright} 2016 The Authors.",
year = "2016",
month = feb,
day = "16",
doi = "10.1016/j.celrep.2016.01.034",
language = "English",
volume = "14",
pages = "1448--1461",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "6",
}
