The p53 tumor suppressor plays a key role in maintaining genomic integrity and preventing tumorigenesis. MDM2, a RING-finger ubiquitin E3 ligase, plays a major role in the regulation of p53 levels. MDM2 ubiquitinates p53 and targets it for proteasome-mediated degradation. MDMX, an MDM2 homolog, also plays an indispensible role in controlling p53. MDMX assists MDM2 in suppressing p53 activity and levels, while MDM2 mediates MDMX ubiquitination and degradation in response to DNA damage. As MDM2, and possibly MDMX as well, can be transcription ally induced by p53, they form a p53-MDM2-MDMX feedback loop. This loop ensues a normal homeostasis of these proteins in cells. Recent studies have also shown that the p53-MDM2-MDMX loop is regulated via deubiquitination by counteraction of several deubiquitinating enzymes (DUBs) from the ubiquitin specific protease (USP) family. USP7 deubiquitinates and stabilizes all the three players, whereas USP10 deubiquitinates p53 only and USP2a deubiquitinates MDM2 and MDMX, providing another layer of precise and dynamic regulation of the p53 pathway. However, whether this loop is regulated by DUBs other than USP family is not known. We recently discovered that an OTU-domain containing protease (OTU) family member, Otubain 1 (Otub1), as a novel regulator of the p53-MDM2-MDMX loop. We show that over expression of Otub1 reverses MDM2-mediated p53 ubiquitination, stabilize p53 in cells, and drastically induce p53- dependent apoptosis and cell growth inhibition. Over expression of a dominant-negative mutant of Otub1 or ablation of endogenous Otub1 by siRNA significantly attenuated p53 activation in response to DNA damage. These results suggest a crucial role for Otub1 in the regulation of p53 signaling. To further gain insight into the regulation, we will investigate the molecular and biochemical mechanisms underlying the role of Otub1 in regulating the p53 pathway in Aim 1. As Otub1 also suppresses MDMX ubiquitination and stabilizes MDMX in cells and, intriguingly, the stabilized MDMX by Otub1 is highly phosphorylated, we will further characterize the mechanism underlying Otub1 regulation of MDMX as well as its significance in Aim 2. Finally, as Otub1 drastically induces p53-dependent cell growth inhibition in cell culture system, we will examine if Otub1 suppresses tumor growth in vivo using mouse xenograft and knockout models in Aim 3. We will examine whether Otub1 expression is deregulated and/or the Otub1 gene is mutated in human cancers. We will also examine the mechanism underlying the Otub1 regulation of p53 signaling in response to DNA damage. Completion of these aims would not only further our understanding of how p53 function is properly regulated through dynamic ubiquitination and deubiquitination and how deregulation of this dynamics contributes to tumorigenesis, but also aid the search for novel cancer therapeutics to re-activate p53 in wild-type p53 containing cancers.

Public Health Relevance

The p53 tumor suppressor plays a key role in suppressing tumor formation and growth. This study will elucidate the molecular mechanisms controlling the cellular levels of p53 and will yield crucial insight into how to reactivate p53 in cancer therapy.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA160474-03
Application #
8634742
Study Section
Molecular Oncogenesis Study Section (MONC)
Program Officer
Watson, Joanna M
Project Start
2012-05-01
Project End
2017-03-31
Budget Start
2014-04-01
Budget End
2015-03-31
Support Year
3
Fiscal Year
2014
Total Cost
$308,659
Indirect Cost
$107,384
Name
Oregon Health and Science University
Department
Genetics
Type
Schools of Medicine
DUNS #
096997515
City
Portland
State
OR
Country
United States
Zip Code
97239
Sun, Xiao-Xin; Chen, Yingxiao; Su, Yulong et al. (2018) SUMO protease SENP1 deSUMOylates and stabilizes c-Myc. Proc Natl Acad Sci U S A 115:10983-10988
DeVine, Tiffany; Sears, Rosalie C; Dai, Mu-Shui (2018) The ubiquitin-specific protease USP36 is a conserved histone H2B deubiquitinase. Biochem Biophys Res Commun 495:2363-2368
Sun, Xiao-Xin; Dai, Mu-Shui (2017) p73 to the rescue: Role of RPL26. Oncotarget 8:5641-5642
Van Hook, Kathryn; Wang, Zhiping; Chen, Dexi et al. (2017) ?N-ASPP2, a novel isoform of the ASPP2 tumor suppressor, promotes cellular survival. Biochem Biophys Res Commun 482:1271-1277
Chen, Yingxiao; Wang, Yue-Gang; Li, Yuhuang et al. (2017) Otub1 stabilizes MDMX and promotes its proapoptotic function at the mitochondria. Oncotarget 8:11053-11062
Tian, Qi; Gromov, Pavel; Clement, Joachim H et al. (2017) RHEB1 insufficiency in aged male mice is associated with stress-induced seizures. Geroscience 39:557-570
Baumann, Douglas G; Dai, Mu-Shui; Lu, Hua et al. (2017) GFZF, a glutathione S-transferase protein implicated in cell cycle regulation and hybrid inviability, is a transcriptional co-activator. Mol Cell Biol :
He, Xia; Li, Yuhuang; Dai, Mu-Shui et al. (2016) Ribosomal protein L4 is a novel regulator of the MDM2-p53 loop. Oncotarget 7:16217-26
Sun, Xiao-Xin; Sears, Rosalie C; Dai, Mu-Shui (2015) Deubiquitinating c-Myc: USP36 steps up in the nucleolus. Cell Cycle 14:3786-93
Lo, D; Zhang, Y; Dai, M-S et al. (2015) Nucleostemin stabilizes ARF by inhibiting the ubiquitin ligase ULF. Oncogene 34:1688-97

Showing the most recent 10 out of 19 publications