KRAS Engages AGO2 to Enhance Cellular Transformation

Sunita Shankar; Sethuramasundaram Pitchiaya; Rohit Malik; Vishal Kothari; Yasuyuki Hosono; Anastasia K Yocum; Harika Gundlapalli; Yasmine White; Ari Firestone; Xuhong Cao; Saravana M Dhanasekaran; Jeanne A Stuckey; Gideon Bollag; Kevin Shannon; Nils G Walter; Chandan Kumar-Sinha; Arul M Chinnaiyan

doi:10.1016/j.celrep.2016.01.034

KRAS Engages AGO2 to Enhance Cellular Transformation

Cell Rep. 2016 Feb 16;14(6):1448-1461. doi: 10.1016/j.celrep.2016.01.034. Epub 2016 Feb 4.

Authors

Sunita Shankar¹, Sethuramasundaram Pitchiaya², Rohit Malik¹, Vishal Kothari¹, Yasuyuki Hosono¹, Anastasia K Yocum¹, Harika Gundlapalli¹, Yasmine White³, Ari Firestone³, Xuhong Cao⁴, Saravana M Dhanasekaran¹, Jeanne A Stuckey⁵, Gideon Bollag⁶, Kevin Shannon³, Nils G Walter⁷, Chandan Kumar-Sinha¹, Arul M Chinnaiyan⁸

Affiliations

¹ Michigan Center for Translational Pathology, University of Michigan, Ann Arbor, MI 48109, USA; Department of Pathology, University of Michigan, Ann Arbor, MI 48109, USA.
² Michigan Center for Translational Pathology, University of Michigan, Ann Arbor, MI 48109, USA; Department of Pathology, University of Michigan, Ann Arbor, MI 48109, USA; Single Molecule Analysis Group, Department of Chemistry, University of Michigan, Ann Arbor, MI 48109, USA.
³ Department of Pediatrics and Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, CA 94158, USA.
⁴ Michigan Center for Translational Pathology, University of Michigan, Ann Arbor, MI 48109, USA; Howard Hughes Medical Institute, University of Michigan, Ann Arbor, MI 48109, USA.
⁵ Life Science Institute, University of Michigan, Ann Arbor, MI 48109, USA; Department of Biological Chemistry, University of Michigan, Ann Arbor, MI 48109, USA.
⁶ Plexxikon Inc., Berkeley, CA 94710, USA.
⁷ Single Molecule Analysis Group, Department of Chemistry, University of Michigan, Ann Arbor, MI 48109, USA.
⁸ Michigan Center for Translational Pathology, University of Michigan, Ann Arbor, MI 48109, USA; Department of Pathology, University of Michigan, Ann Arbor, MI 48109, USA; Howard Hughes Medical Institute, University of Michigan, Ann Arbor, MI 48109, USA; Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI 48109, USA; Department of Urology, University of Michigan, Ann Arbor, MI 48109, USA. Electronic address: arul@umich.edu.

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 KRAS(G12V)-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; EIF2C2; KRAS; RNA silencing; cancer.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Argonaute Proteins / antagonists & inhibitors
Argonaute Proteins / genetics*
Argonaute Proteins / metabolism
Base Sequence
Carboxypeptidases / genetics
Carboxypeptidases / metabolism
Cell Transformation, Neoplastic / genetics*
Cell Transformation, Neoplastic / metabolism
Cell Transformation, Neoplastic / pathology
Endoplasmic Reticulum / metabolism*
Gene Expression Regulation, Neoplastic*
Gene Silencing
Humans
Mice
MicroRNAs / genetics
MicroRNAs / metabolism
Molecular Sequence Data
Mutation
NIH 3T3 Cells
Phosphatidylinositol 3-Kinases / genetics
Phosphatidylinositol 3-Kinases / metabolism
Protein Binding
Proto-Oncogene Proteins p21(ras) / genetics*
Proto-Oncogene Proteins p21(ras) / metabolism
RNA, Small Interfering / genetics
RNA, Small Interfering / metabolism
Transgenes

Substances

AGO2 protein, human
Argonaute Proteins
KRAS protein, human
MicroRNAs
RNA, Small Interfering
Phosphatidylinositol 3-Kinases
Carboxypeptidases
SCPEP1 protein, human
Proto-Oncogene Proteins p21(ras)

Abstract

Publication types

MeSH terms

Substances

Grants and funding