Utilization of Oriented Peptide Libraries to Identify Substrate Motifs Selected by ATM

O'Neill, Ted B.; Dwyer, Alison; Ziv, Yael; Chan, Doug W.; Lees‐Miller, Susan P.; Abraham, Robert; Lai, Jack H.; Hill, David R.C.; Shiloh, Yossi; Cantley, Lewis C.; Rathbun, Gary

doi:10.1074/jbc.m001002200

Cited by 176 publications

(133 citation statements)

References 62 publications

(63 reference statements)

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“…The carboxy-terminal domain of HMGA2 harbors a SQ motif (O'Neill et al, 2000) (serine 102/glutamine 103), which corresponds to the serine 88/glutamine 89 of HMGA1, phosphorylated by ATM (Pentimalli et al, 2008). Therefore, we investigated whether HMGA2 is also a novel target of ATM kinase activity (Figure1 and Supplementary Figure S1).…”

Section: Hmga2 Interacts With and Is Phosphorylated By Atmmentioning

confidence: 99%

HMGA proteins promote ATM expression and enhance cancer cell resistance to genotoxic agents

et al. 2011

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DNA-damaging therapies represent a keystone in cancer treatment. Unfortunately, many tumors often relapse because of a group of cancer cells, which are resistant to conventional therapies. High-mobility group A (HMGA) proteins has a key role in cell transformation, and their overexpression is a common feature of human malignant neoplasias, representing a poor prognostic index often correlated to anti-cancer drug resistance. Our previous results demonstrated that HMGA1 is a substrate of ataxiatelangiectasia mutated (ATM), the main cellular sensor of genotoxic stress. Here we also report thatHMGA2, the other member of the HMGA family, is a novel substrate of ATM. Interestingly, we found that HMGA proteins positively regulate ATM gene expression. Moreover, induction of ATM kinase activity by DNA-damaging agents enhances HMGA-dependent transcriptional activation of ATM promoter, suggesting that ATM expression is modulated by a DNA-damage-and HMGA-dependent positive feedback loop. Finally, inhibition of HMGA expression in mouse embryonic fibroblasts and in cancer cells strongly reduces ATM protein levels, impairing the cellular DNA-damage response and enhancing the sensitivity to DNA-damaging agents. These findings indicate this novel HMGA-ATM pathway as a new potential target to improve the effectiveness of conventional anti-neoplastic treatments on the genotoxic-drug resistant cancer cells.

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Section: Hmga2 Interacts With and Is Phosphorylated By Atmmentioning

confidence: 99%

HMGA proteins promote ATM expression and enhance cancer cell resistance to genotoxic agents

et al. 2011

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“…Previous studies have found that FUS is phosphorylated by phosphoinositide 3‐kinase‐like kinases (PIKKs), especially DNA‐dependent protein kinase (DNA‐PK; Gardiner et al , 2008; Deng et al , 2014). PIKKs phosphorylate their protein targets at serine or threonine residues followed by glutamine (S/TQ; Kim et al , 1999; O'Neill et al , 2000). Indeed, the FUS LC domain has twelve S/TQ motifs (8 SQ, 4 TQ), but previous in vitro DNA‐PK phosphorylation of FUS identified only pS26, pS42, pS61, pS84, and pS131 (Gardiner et al , 2008; Han et al , 2012).…”

Section: Introductionmentioning

confidence: 99%

Phosphorylation of the FUS low‐complexity domain disrupts phase separation, aggregation, and toxicity

et al. 2017

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Neuronal inclusions of aggregated RNA‐binding protein fused in sarcoma (FUS) are hallmarks of ALS and frontotemporal dementia subtypes. Intriguingly, FUS's nearly uncharged, aggregation‐prone, yeast prion‐like, low sequence‐complexity domain (LC) is known to be targeted for phosphorylation. Here we map in vitro and in‐cell phosphorylation sites across FUS LC. We show that both phosphorylation and phosphomimetic variants reduce its aggregation‐prone/prion‐like character, disrupting FUS phase separation in the presence of RNA or salt and reducing FUS propensity to aggregate. Nuclear magnetic resonance spectroscopy demonstrates the intrinsically disordered structure of FUS LC is preserved after phosphorylation; however, transient domain collapse and self‐interaction are reduced by phosphomimetics. Moreover, we show that phosphomimetic FUS reduces aggregation in human and yeast cell models, and can ameliorate FUS‐associated cytotoxicity. Hence, post‐translational modification may be a mechanism by which cells control physiological assembly and prevent pathological protein aggregation, suggesting a potential treatment pathway amenable to pharmacologic modulation.

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“…The most stringent classification requires a minimum of five SQ/TQ sites within a span of 50 residues with preferred phosphorylation sites generally preceded by a hydrophobic residue such as leucine (40,41). The Sp1 sequence spanning residues 56 to 102 easily fulfills this criterion, containing five SQ/TQ sites within 47 amino acids (Fig.…”

Section: Sp1 Is Phosphorylated In Response To H 2 Omentioning

confidence: 99%

Phosphorylation of Sp1 in Response to DNA Damage by Ataxia Telangiectasia-Mutated Kinase

Olofsson

Kelly

Kim

et al. 2007

Molecular Cancer Research

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Sp1, a transcription factor that regulates expression of a wide array of essential genes, contains two SQ/TQ cluster domains, which are characteristic of ATM kinase substrates. ATM substrates are transducers and effectors of the DNA damage response, which involves sensing damage, checkpoint activation, DNA repair, and/or apoptosis. A role for Sp1 in the DNA damage response is supported by our findings: Activation of ATM induces Sp1 phosphorylation with kinetics similar to H2AX; inhibition of ATM activity blocks Sp1 phosphorylation; depletion of Sp1 sensitizes cells to DNA damage and increases the frequency of double strand breaks. We have identified serine 101 as a critical site phosphorylated by ATM; Sp1 with serine 101 mutated to alanine (S101A) is not significantly phosphorylated in response to damage and cannot restore increased sensitivity to DNA damage of cells depleted of Sp1. Together, these data show that Sp1 is a novel ATM substrate that plays a role in the cellular response to DNA damage. (Mol Cancer Res 2007;5(12):1319 -30)

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Utilization of Oriented Peptide Libraries to Identify Substrate Motifs Selected by ATM

Cited by 176 publications

References 62 publications

HMGA proteins promote ATM expression and enhance cancer cell resistance to genotoxic agents

HMGA proteins promote ATM expression and enhance cancer cell resistance to genotoxic agents

Phosphorylation of the FUS low‐complexity domain disrupts phase separation, aggregation, and toxicity

Phosphorylation of Sp1 in Response to DNA Damage by Ataxia Telangiectasia-Mutated Kinase

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