Gene Synthesis, Expression, and Processing of Human Ubiquitin Carboxyl Extension Proteins

Monia, Brett P.; Ecker, David J.; Jonnalagadda, Sobhanaditya; Marsh, Jeanne C.; Gotlib, Leah; Butt, Tauseef R.; Crooke, Stanley T.

doi:10.1016/s0021-9258(19)84967-0

Cited by 57 publications

(11 citation statements)

References 34 publications

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“…RPS27a is overexpressed in renal, breast and colon carcinomas [ 54 , 55 ], and its gene expression has been found to be markedly elevated in an oncomouse model of hepatocellular carcinoma [ 56 ]. It also has an essential role in the activation of cellular checkpoints via p53 [ 57 ].…”

Section: Discussionmentioning

confidence: 99%

Loss of RPS27a expression regulates the cell cycle, apoptosis, and proliferation via the RPL11-MDM2-p53 pathway in lung adenocarcinoma cells

Zhang

Huang

et al. 2022

J Exp Clin Cancer Res

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Background Depletion of certain ribosomal proteins induces p53 activation, which is mediated mainly by ribosomal protein L5 (RPL5) and/or ribosomal protein L11 (RPL11). Therefore, RPL5 and RPL11 may link RPs and p53 activation. Thus, this study aimed to explore whether RPs interact with RPL11 and regulate p53 activation in lung adenocarcinoma (LUAD) cells. Methods The endogenous RPL11-binding proteins in A549 cells were pulled down through immunoprecipitation and identified with a proteomics approach. Docking analysis and GST-fusion protein assays were used to analyze the interaction of ribosomal protein S27a (RPS27a) and RPL11. Co-immunoprecipitation and in vitro ubiquitination assays were used to detect the effects of knockdown of RPS27a on the interaction between RPS27a and RPL11, and on p53 accumulation. Cell cycle, apoptosis, cell invasion and migration, cell viability and colony-formation assays were performed in the presence of knockdown of RPS27a. The RPS27a mRNA expression in LUAD was analyzed on the basis of the TCGA dataset, and RPS27a expression was detected through immunohistochemistry in LUAD samples. Finally, RPS27a and p53 expression was analyzed through immunohistochemistry in A549 cell xenografts with knockdown of RPS27a. Results RPS27a was identified as a novel RPL11 binding protein. GST pull-down assays revealed that RPS27a directly bound RPL11. Knockdown of RPS27a weakened the interaction between RPS27a and RPL11, but enhanced the binding of RPL11 and murine double minute 2 (MDM2), thereby inhibiting the ubiquitination and degradation of p53 by MDM2. Knockdown of RPS27a stabilized p53 in an RPL11-dependent manner and induced cell viability inhibition, cell cycle arrest and apoptosis in a p53-dependent manner in A549 cells. The expression of RPS27a was upregulated in LUAD and correlated with LUAD progression and poorer prognosis. Overexpression of RPS27a correlated with upregulation of p53, MDM2 and RPL11 in LUAD clinical specimens. Knockdown of RPS27a increased p53 activation, thus, suppressing the formation of A549 cell xenografts in nude mice. Conclusions RPS27a interacts with RPL11, and RPS27a knockdown enhanced the binding of RPL11 and MDM2, thereby inhibiting MDM2-mediated p53 ubiquitination and degradation; in addition, RPS27a as important roles in LUAD progression and prognosis, and may be a therapeutic target for patients with LUAD. Graphical Abstract

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Section: Discussionmentioning

confidence: 99%

Loss of RPS27a expression regulates the cell cycle, apoptosis, and proliferation via the RPL11-MDM2-p53 pathway in lung adenocarcinoma cells

Zhang

Huang

et al. 2022

J Exp Clin Cancer Res

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“…9) seem to be identical to those of ubiquitination. Ubiquitin is expressed as a polyubiquitin moiety or as an NH 2 -terminal fusion to unrelated proteins and must be proteolytically processed to release monomeric ubiquitin and to reveal the COOH-terminal Gly-Gly motif that is involved in coupling to targets (e.g., Ozkaynak et al, 1984;Monia et al, 1989;Callis et al, 1995). In comparison, SUMO-1 is expressed as a single 101-amino acid protein that contains 4 additional COOH-terminal amino acids beyond the Gly-Gly motif that it shares with ubiquitin (Mahajan et al, 1997).…”

Section: Identification Of the Rangap1-sumo-1 Link Reveals Similarity To Ubiquitinationmentioning

confidence: 99%

Molecular Characterization of the SUMO-1 Modification of RanGAP1 and Its Role in Nuclear Envelope Association

Mahajan

Gerace

Melchior

1998

The Journal of Cell Biology

265

230

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The mammalian guanosine triphosphate (GTP)ase-activating protein RanGAP1 is the first example of a protein covalently linked to the ubiquitin-related protein SUMO-1. Here we used peptide mapping, mass spectroscopy analysis, and mutagenesis to identify the nature of the link between RanGAP1 and SUMO-1. SUMO-1 is linked to RanGAP1 via glycine 97, indicating that the last 4 amino acids of this 101– amino acid protein are proteolytically removed before its attachment to RanGAP1. Recombinant SUMO-1 lacking the last four amino acids is efficiently used for modification of RanGAP1 in vitro and of multiple unknown proteins in vivo. In contrast to most ubiquitinated proteins, only a single lysine residue (K526) in RanGAP1 can serve as the acceptor site for modification by SUMO-1. Modification of RanGAP1 with SUMO-1 leads to association of RanGAP1 with the nuclear envelope (NE), where it was previously shown to be required for nuclear protein import. Sufficient information for modification and targeting resides in a 25-kD domain of RanGAP1. RanGAP1–SUMO-1 remains stably associated with the NE during many cycles of in vitro import. This indicates that removal of RanGAP1 from the NE is not a required element of nuclear protein import and suggests that the reversible modification of RanGAP1 may have a regulatory role.

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“…The low-expressed RPS27a may have the aforementioned function. RPS27a have been found overexpressed in renal, breast and colon carcinomas [41,42], a dramatic increase in the expression of RPS27a gene in an oncomouse model of hepatocellular carcinoma [43], it is essential role in the activation of cellular checkpoints via p53.…”

Section: Discussionmentioning

confidence: 99%

Loss of RPS27a expression regulates cell cycle, apoptosis, and proliferation via the RPS27a-RPL11-MDM2-p53 pathway in lung adenocarcinoma cells

Zhang

Huang

et al. 2021

Preprint

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Background Some ribosomal proteins (RPs) might regulate the MDM2–p53 loop by binding to RPL5 or RPL11. This study aimed to explore whether ribosomal protein S27a (RPS27a) interacted with the ribosomal protein L11 (RPL11) to regulate p53 in lung adenocarcinoma (LUAD) cells. Methods RPL11-interacting proteins were identified using a proteomics approach. Co-immunoprecipitation (co-IP), docking analysis, GST-fusion and in vitro ubiquitination assay were used to analyze the interaction of RPS27a and RPL11. Cell cycle, apoptosis, cell invasion, cell viability and colony-formation assay were analyzed by knocking down RPS27a. The RPS27a mRNA expression in LUAD was analyzed based on the TCGA dataset and the RPS27a expression was detected by immunohistochemistry in LUAD samples. At last, the RPS27a and p53 expression were analyzed by immunohistochemistry in xenograft tumors by blocking RPS27a. Results The ablation of RPS27a inhibits murine double minute 2 (MDM2)-mediated p53 ubiquitination, induced G1/S cell cycle arrest and apoptosis, and inhibited the proliferation of LUAD cells. Also, it induced p53-dependent cell cycle arrest and RPL11-dependent p53 activation. The protein–protein docking results revealed that RPS27a and RPL11 formed a stable complex structure. The GST-fusion protein–protein association assay demonstrated that RPS27a bound to RPL11. The overexpressed RPS27a in LUAD was found to be correlated with a poorer prognosis based on the TCGA dataset. RPS27a expression was high in patients with LUAD. Blocking RPS27a increased p53, thus, suppressing cell proliferation and A549 xenograft growth in nude mice. Conclusions This study was novel in reporting that RPs bound to RPL11 to regulate the MDM2-p53 feedback loop, revealing that RPS27a plays an important function in LUAD growth. Hence, RPS27a might provide a diagnostic marker or therapeutic target for patients with LUAD.

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Gene Synthesis, Expression, and Processing of Human Ubiquitin Carboxyl Extension Proteins

Cited by 57 publications

References 34 publications

Loss of RPS27a expression regulates the cell cycle, apoptosis, and proliferation via the RPL11-MDM2-p53 pathway in lung adenocarcinoma cells

Loss of RPS27a expression regulates the cell cycle, apoptosis, and proliferation via the RPL11-MDM2-p53 pathway in lung adenocarcinoma cells

Molecular Characterization of the SUMO-1 Modification of RanGAP1 and Its Role in Nuclear Envelope Association

Loss of RPS27a expression regulates cell cycle, apoptosis, and proliferation via the RPS27a-RPL11-MDM2-p53 pathway in lung adenocarcinoma cells

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