The mammalian target of rapamycin complex 1 (mTORC1) is an evolutionarily conserved multi- protein kinase complex that couples nutrient and growth factor sensing to regulate a wide array of essential cellular processes. mTORC1 stimulates protein synthesis and ribosome biogenesis, which are vital cellular processes that regulate cell growth and proliferation. Hyperactivation of mTORC1 signaling is found in many types of cancer cells and leads to aberrant increases in protein synthesis and uncontrolled cell proliferation. Despite the positive roles of mTORC1 in enhancing general cap- dependent translation through eIF4E activation, recent studies reveal mTORC1 preferentially stimulates a subset of mRNAs, including those that encode proteins required for translation, and cell growth, proliferation, and invasion. Interestingly, these mRNAs possess pyrimidine-enriched sequences (PES) such as 5'TOP in their 5'UTRs that are thought to play important roles in their expression. However, the molecular mechanisms by which mTORC1 primarily facilitates the translation of a subset of those mRNAs remain elusive. We have discovered a novel mTORC1 substrate, LARP1 that directly interacts with the PES in the 5'UTRs of 5' TOP mRNAs. Our preliminary data suggest that LARP1 functions not only as a molecular switch to regulate PES- containing mRNA translation in response to mTOR activity, but also as a regulator for mTORC1 to phosphorylate 4EBP1 and S6K1 on translationally-competent mRNAs. Knockdown of LARP1 significantly reduces translation of mRNAs that interact with LARP1 and dramatically inhibits ribosome biogenesis and cell proliferation in various cancer cells. We propose that LARP1 is an atypical mTORC1 substrate that regulates the translation of specific PES mRNAs and enhances their mTORC1-dependent translation. We will explore the molecular mechanisms by which mTORC1- dependent phosphorylation of LARP1 contributes to the translation of its interacting mRNAs. Second, we will determine the functional importance of the 5'UTR PES motif enriched in LARP1-bound mRNAs. Finally, we will explore the in vivo functions of LARP1 as well as pathophysiological role of LARP1 in PI3K-Akt-mTORC1-mediated cancer development in animal models. We will accomplish our goals by exploiting a multifaceted but integrated approach combining biochemistry, molecular biology, proteomics, high-throughput RNA sequencing, and mouse models. By elucidating the molecular mechanisms of mTORC1-LARP1-depenent translation, our proposed studies will provide new insights into the regulations of mTORC1-mediated translation and lead to new therapeutic approaches for cancer that target downstream effectors of mTORC1 activity.

Public Health Relevance

Hyper-activation of mammalian target of rapamycin (mTOR) signaling is often found in a wide array of cancers. mTOR inhibitors are promising anti-cancer drugs and several are currently used or under clinical trials. Dysregulation of mTOR signaling leads to increased synthesis of translational machinery, and subsequently to global protein synthesis that is critical for cancer cell proliferation, invasion, and migration. However, the mechanisms by which mTOR stimulates the production of such specific proteins are not well understood. In this proposal, we will investigate a novel mTOR substrate that plays a key role in mTOR-medicated protein synthesis. Completion of this project will provide crucial information by which the mTOR signaling contributes to cell growth, proliferation, cancer development, and sets the stage for developing new therapeutic approaches targeting novel mTORC1 substrates for cancer treatment.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM110019-04
Application #
9394026
Study Section
Molecular and Integrative Signal Transduction Study Section (MIST)
Program Officer
Koduri, Sailaja
Project Start
2015-04-15
Project End
2018-12-31
Budget Start
2018-01-01
Budget End
2018-12-31
Support Year
4
Fiscal Year
2018
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Physiology
Type
Schools of Medicine
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Hong, Sungki; Freeberg, Mallory A; Han, Ting et al. (2017) LARP1 functions as a molecular switch for mTORC1-mediated translation of an essential class of mRNAs. Elife 6:
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