Abstract
Mono(ADP-ribosyl)ation (MARylation) is emerging as a critical regulator of ribosome function and translation. Herein, we demonstrate that RACK1, an integral component of the ribosome, is MARylated on three acidic residues by the mono(ADP-ribosyl) transferase (MART) PARP14 in ovarian cancer cells. MARylation of RACK1 is required for stress granule formation and promotes the colocalization of RACK1 in stress granules with G3BP1, eIF3η, and 40S ribosomal proteins. In parallel, we observed reduced translation of a subset of mRNAs, including those encoding key cancer regulators (e.g., AKT). Treatment with a PARP14 inhibitor or mutation of the sites of MARylation on RACK1 blocks these outcomes, as well as the growth of ovarian cancer cells in culture and in vivo. To re-set the system after prolonged stress and recovery, the ADP-ribosyl hydrolase TARG1 deMARylates RACK1, leading to the dissociation of the stress granules and the restoration of translation. Collectively, our results demonstrate a therapeutically targetable pathway that controls stress granule assembly and disassembly in ovarian cancer cells.
Summary We have discovered a druggable PARP14/TARG1-regulated pathway that mediates site- specific mono(ADP-ribosyl)ation of RACK1, a ribosomal protein. This pathway controls stress granule assembly and disassembly, as well as the translation of a subset of mRNAs, to modulate the growth of ovarian cancer cells in culture and in vivo.
Competing Interest Statement
W.L.K. is a founder, member of the SAB, member of the BOD, and a stockholder for and ARase Therapeutics, Inc. He is also coholder of U.S. Patent 9,599,606 covering the ADP-ribose detection reagents used herein, which has been licensed to and is sold by EMD Millipore.
Footnotes
↵8 Lead Contact / Address manuscript correspondence to: W. Lee Kraus, Ph.D. Cecil H. and Ida Green Center for Reproductive Biology Sciences The University of Texas Southwestern Medical Center at Dallas 5323 Harry Hines Boulevard Dallas, TX 75390-8511 Phone: 214-648-2388 Fax: 214-648-0383 E-mail: LEE.KRAUS{at}utsouthwestern.edu
Conflict of Interest Statement: W.L.K. is a founder, member of the SAB, member of the BOD, and a stockholder for ARase Therapeutics, Inc. He is also coholder of U.S. Patent 9,599,606 covering the ADP-ribose detection reagents used herein, which has been licensed to and is sold by EMD Millipore.
We have addressed the comments of three reviewers for Journal of Cell Biology to solidify our results and add support for our conclusions.
