Hyperactivation of EGFR and downstream effector phospholipase D1 by oncogenic FAM83B

Cipriano, Rocky; Bryson, Benjamin L.; Miskimen, Kristy; Bartel, Courtney A.; Hernandez-Sanchez, Wilnelly; Bruntz, Ronald C.; Scott, Sarah; Lindsley, Craig W.; Brown, H. Alex; Jackson, Mark W.

doi:10.1038/onc.2013.293

Cited by 36 publications

(34 citation statements)

References 38 publications

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“…Given the recent finding that elevated ERK activation in basal-like breast cancer conferred elevated proliferation rates even following neoadjuvant therapy (14), we propose that the inhibition of key intermediaries involved in MAPK activation, such as the FAM83 proteins, may facilitate genotoxic killing of these difficult to treat cancers. In addition, we recently identified that elevated FAM83B expression also activates the PI3K/AKT signaling pathway and EGFR/PLD1 axis (15, 16). The combined activation of both MAPK and PI3K/AKT signaling resulted in a decreased sensitivity to numerous targeted therapies, including EGFR, PI3K, AKT, and mTOR inhibitors.…”

Section: Discussionmentioning

confidence: 99%

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Conserved Oncogenic Behavior of the FAM83 Family Regulates MAPK Signaling in Human Cancer

Cipriano¹,

Miskimen²,

Bryson³

et al. 2014

Molecular Cancer Research

109

103

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FAM83B (Family with sequence similarity 83, member B) was recently identified as a novel oncogene involved in activating CRAF/MAPK signaling and driving epithelial cell transformation. FAM83B is one of eight members of a protein family (FAM83) characterized by a highly conserved domain of unknown function (DUF1669), which is necessary and sufficient to drive transformation. Here, it is demonstrated that additional FAM83 members also exhibit oncogenic properties and have significantly elevated levels of expression in multiple human tumor types using a TissueScan Cancer Survey Panel PCR array and database mining. Furthermore, modeling the observed tumor expression of FAM83A, FAM83C, FAM83D, or FAM83E promoted human mammary epithelial cell (HMEC) transformation, which correlated with the ability of each FAM83 member to bind CRAF (RAF1) and promote CRAF membrane localization. Conversely, ablation of FAM83A or FAM83D from breast cancer cells resulted in diminished MAPK signaling with marked suppression of growth in vitro and tumorigenicity in vivo. Importantly, each FAM83 member was determined to be elevated in at least one of 17 distinct tumor types examined, with FAM83A, FAM83B, and FAM83D most frequently overexpressed in several diverse tissue types. Finally, evidence suggests that elevated expression of FAM83 members is associated with tumor grade and overall survival. Implications FAM83 proteins represent a novel family of oncogenes suitable for the development of cancer therapies aimed at suppressing MAPK signaling.

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Section: Discussionmentioning

confidence: 99%

“…The DUF1669 of FAM83B is sufficient to drive transformation, and interact with CRAF and EGFR (6, 15). Furthermore, FAM83A is the smallest FAM83 member at 434 amino acids, and is comprised mainly of the DUF1669.…”

Section: Discussionmentioning

confidence: 99%

Conserved Oncogenic Behavior of the FAM83 Family Regulates MAPK Signaling in Human Cancer

Cipriano¹,

Miskimen²,

Bryson³

et al. 2014

Molecular Cancer Research

109

103

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“…Although many of these putative SMYD2 substrates are largely functionally uncharacterized, there are nonetheless several substrates that raise intriguing possibilities for potential roles for SMYD2 in progrowth signaling pathways. For example, there are now multiple lines of evidence supporting an oncogenic role for FAM83B, in which we identified SMYD2-mediated monomethylation sites at K652 and K661 (51)(52)(53). We also identified a SMYD2-mediated mono-methylation site at K1304 in RICTOR, a component of the mTORC2 kinase complex (54) as well as in the intracellular domain of GPR126, an adhesion G-protein coupled receptor that bridges type-IV collagen in the extracellular matrix to intracellular cyclic AMP signaling (55).…”

Section: Discussionmentioning

confidence: 99%

Quantitative Profiling of the Activity of Protein Lysine Methyltransferase SMYD2 Using SILAC-Based Proteomics

Olsen

Cao

Han

et al. 2016

Molecular & Cellular Proteomics

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The significance of non-histone lysine methylation in cell biology and human disease is an emerging area of research exploration. The development of small molecule inhibitors that selectively and potently target enzymes that catalyze the addition of methyl-groups to lysine residues, such as the protein lysine mono-methyltransferase SMYD2, is an active area of drug discovery. Critical to the accurate assessment of biological function is the ability to identify target enzyme substrates and to define enzyme substrate specificity within the context of the cell. Here, using stable isotopic labeling with amino acids in cell culture (SILAC) coupled with immunoaffinity enrichment of mono-methyl-lysine (Kme1) peptides and mass spectrometry, we report a comprehensive, large-scale proteomic study of lysine mono-methylation, comprising a total of 1032 Kme1 sites in esophageal squamous cell carcinoma (ESCC) cells and 1861 Kme1 sites in ESCC cells overexpressing SMYD2. Among these Kme1 sites is a subset of 35 found to be potently down-regulated by both shRNA-mediated knockdown of SMYD2 and LLY-507, a selective small molecule inhibitor of SMYD2. In addition, we report specific protein sequence motifs enriched in Kme1 sites that are directly regulated by endogenous SMYD2 activity, revealing that SMYD2 substrate specificity is more diverse than expected. We further show direct activity of SMYD2 toward BTF3-K2, PDAP1-K126 as well as numerous sites within the repetitive units of two unique and exceptionally large proteins, AHNAK and AHNAK2. Collectively, our findings provide quantitative insights into the cellular activity and substrate recognition of SMYD2 as well as the global landscape and regulation of protein mono-methylation. Protein lysine methyltransferases (PKMTs) 1 catalyze the sequence-specific transfer of one, two, or three methyl groups to the side chains of lysine residues (1-4). In addition to the extensively studied lysine methylation on histones, PKMTs can modify non-histone proteins (3,(5)(6)(7)(8)). An increasing number of non-histone proteins have been reported as PKMT substrates, and, as a result, potential roles for the dysregulation of non-histone lysine methylation in cancer development and progression have been proposed (9). The majority of PKMT substrates have been identified based on biochemical methylation assays using recombinant enzyme and substrate, followed by cell-based assays typically requiring overexpression of enzyme and/or substrate to maximize signal detection (10 -13). However, it is difficult to discern whether these substrates are bona fide physiological substrates of endogenous PKMT activity. With the development of PKMT-targeted drugs emerging as a key area of drug discovery (14 -18), unbiased and quantitative methods enabling the comprehensive identification of histone and non-histone substrates in cells are critical to clarifying the cell-relevant substrates of these enzymes and to more accurately understand the functions of non-histone methylation.Progressing from targeted biochemical...

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“…EGFR directly interacts with phospholipase D2 (PLD2) (4-6). Stimulation of EGFR increases cellular PLD activity and the production of phosphatidic acid (PA) in cancer cell lines (7,8). PA is also the precursor to lysophosphatidic acid (LPA) that is relevant in ovarian cancer (9).…”

mentioning

confidence: 99%

Phosphatidic Acid Increases Epidermal Growth Factor Receptor Expression by Stabilizing mRNA Decay and by Inhibiting Lysosomal and Proteasomal Degradation of the Internalized Receptor

Hatton

Lintz

Mahankali

et al. 2015

Molecular and Cellular Biology

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Overexpression of epidermal growth factor receptor (EGFR) is one of the frequent mechanisms implicated in cancer progression, and so is the overexpression of the enzyme phospholipase D (PLD) and its reaction product, phosphatidic acid (PA). However, an understanding of how these signaling molecules interact at the level of gene expression is lacking. Catalytically active PLD enhanced expression of EGFR in human breast cancer cells. Overexpression of the PLD2 isoform increased EGFR mRNA and protein expression. It also negated an EGFR downregulation mediated by small interfering RNA targeting EGFR (siEGFR). Several mechanisms contributed to the alteration in EGFR expression. First was the stabilization of EGFR transcripts as PLD2 delayed mRNA decay, which prolonged their half-lives. Second, RNase enzymatic activity was inhibited by PA. Third, protein stabilization also occurred, as indicated by PLD resistance to cycloheximide-induced EGFR protein degradation. Fourth, PA inhibited lysosomal and proteasomal degradation of internalized EGFR. PLD2 and EGFR colocalized at the cell membrane, and JAK3 phosphorylation at Tyr980/Tyr981 followed receptor endocytosis. Further, the presence of PLD2 increased stabilization of intracellular EGFR in large recycling vesicles at ϳ15 min of EGF stimulation. Thus, PLD2-mediated production of PA contributed to the control of EGFR exposure to ligand through a multipronged transcriptional and posttranscriptional program during the out-of-control accumulation of EGFR signaling in cancer cells. Epidermal growth factor receptor (EGFR) is overexpressed in many epithelial tumors, including bladder, kidney, pancreatic, and squamous cell carcinomas (1). In breast cancer, EGFR overexpression is associated with advanced-stage disease and shortened relapse-free survival, which occurs concomitantly with low estrogen receptor expression (2). The cell signaling events after EGFR ligand stimulation and cancer have been greatly studied and require association of the receptor with a number of cytoplasmic tyrosine kinases, as well as activation of the Janus kinase (JAK)/ STAT pathway (3). EGFR directly interacts with phospholipase D2 (PLD2) (4-6). Stimulation of EGFR increases cellular PLD activity and the production of phosphatidic acid (PA) in cancer cell lines (7,8). PA is also the precursor to lysophosphatidic acid (LPA) that is relevant in ovarian cancer (9). PLD2 activity is regulated by the phosphorylation of the kinases EGFR and Janus kinase 3 (JAK3) on the Y296 and Y415 tyrosine residues, respectively (10, 11). JAK3 is a 130-kDa intracellular, nonreceptor tyrosine kinase (12, 13). It can also function as the docking site for other proteins if they have the proper Src homology 2 (SH2) domains.Tyrosine kinases associated with EGFR, such as Fer/Fes, promote cell motility in a PLD/PA-dependent pathway (14). We have recently found that Fes binds PA and participates in a PLD-induced pathway of myeloid differentiation (15). PLD2 is associated with EGFR signaling by binding to Grb2 at two specific res...

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Hyperactivation of EGFR and downstream effector phospholipase D1 by oncogenic FAM83B

Cited by 36 publications

References 38 publications

Conserved Oncogenic Behavior of the FAM83 Family Regulates MAPK Signaling in Human Cancer

Conserved Oncogenic Behavior of the FAM83 Family Regulates MAPK Signaling in Human Cancer

Quantitative Profiling of the Activity of Protein Lysine Methyltransferase SMYD2 Using SILAC-Based Proteomics

Phosphatidic Acid Increases Epidermal Growth Factor Receptor Expression by Stabilizing mRNA Decay and by Inhibiting Lysosomal and Proteasomal Degradation of the Internalized Receptor

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