Wortmannin, a Widely Used Phosphoinositide 3-Kinase Inhibitor, also Potently Inhibits Mammalian Polo-like Kinase

Liu, Yongsheng; Shreder, Kevin; Gai, Wenzhi; Corral, Sergio; Ferris, Douglas K.; Rosenblum, Jonathan S.

doi:10.1016/j.chembiol.2004.11.009

Cited by 189 publications

(158 citation statements)

References 51 publications

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“…The active site lysine (Lys802 in p110α and Lys833 in p110γ) is critical for catalysis and is highly conserved in lipid and protein kinases. Wortmannin also targets class II PI3K C2β (IC 50 =1.6 nM), the human class III PI3K hVps34p (IC 50 = 2.5 nM), type III PI4Ks (IC 50 =50-300 nM) and polo-like protein kinases that regulate mitosis (IC 50 =24-48 nM) [60][61][62][63][64]. Not surprisingly, considering the similarity between the PI3K and PIKK catalytic domains, wortmannin also irreversibly inhibits PIKK family members.…”

Section: Mtor Kinase Inhibitorsmentioning

confidence: 99%

Rapamycin and mTOR kinase inhibitors

2008

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Mammalian target of rapamycin (mTOR) is a protein kinase that controls cell growth, proliferation, and survival. mTOR signaling is often upregulated in cancer and there is great interest in developing drugs that target this enzyme. Rapamycin and its analogs bind to a domain separate from the catalytic site to block a subset of mTOR functions. These drugs are extremely selective for mTOR and are already in clinical use for treating cancers, but they could potentially activate an mTOR-dependent survival pathway that could lead to treatment failure. By contrast, small molecules that compete with ATP in the catalytic site would inhibit all of the kinase-dependent functions of mTOR without activating the survival pathway. Several non-selective mTOR kinase inhibitors have been described and here we review their chemical and cellular properties. Further development of selective mTOR kinase inhibitors holds the promise of yielding potent anticancer drugs with a novel mechanism of action.

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Section: Mtor Kinase Inhibitorsmentioning

confidence: 99%

Rapamycin and mTOR kinase inhibitors

2008

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“…In Vitro Kinase Reaction-To identify the possible Thr residue(s) of p73␣ that could be phosphorylated by Plk1, we used CycLex Polo-like kinase 1 assay kit (CycLex, Nagano, Japan) (29). In brief, the purified Plk1 was added to the reaction mixture containing 50 M ATP and polyclonal anti-phospho-Thr antibody and incubated for 30 min at room temperature.…”

Section: Construction Of Mutant Forms Of Gst-p73␣-(1-130)-mentioning

confidence: 99%

Inhibitory Role of Plk1 in the Regulation of p73-dependent Apoptosis through Physical Interaction and Phosphorylation

Koida

Ozaki

Yamamoto

et al. 2008

Journal of Biological Chemistry

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In response to DNA damage, p73 plays a critical role in cell fate determination. In this study, we have found that Plk1 (pololike kinase 1) associates with p73, phosphorylates p73 at Thr-27, and thereby inhibits its pro-apoptotic activity. During cisplatinmediated apoptosis in COS7 cells in which the endogenous p53 is inactivated by SV40 large T antigen, p73 was induced to accumulate in association with a significant down-regulation of Plk1. Consistent with these observations, Plk1 reduced the stability of the endogenous p73. Immunoprecipitation and in vitro pulldown assay demonstrated that p73 binds to the kinase domain of Plk1 through its NH 2 -terminal region. Luciferase reporter assay and reverse transcription-PCR analysis revealed that Plk1 is able to block the p73-mediated transcriptional activation. Of note, kinase-deficient Plk1 mutant (Plk1(K82M)) retained an ability to interact with p73; however, it failed to inactivate the p73-mediated transcriptional activation, suggesting that kinase activity of Plk1 is required for the inhibition of p73. Indeed, in vitro kinase assay indicated that p73 is phosphorylated at Thr-27 by Plk1. Furthermore, small interference RNA-mediated knockdown of the endogenous Plk1 in p53-deficient H1299 cells resulted in a significant increase in the number of cells with sub-G 1 DNA content accompanied by the upregulation of p73 and pro-apoptotic p53 AIP1 as well as the proteolytic cleavage of poly(ADP-ribose) polymerase. Thus, our present results suggest that Plk1-mediated dysfunction of p73 is one of the novel molecular mechanisms to inhibit the p53-independent apoptosis, and the inhibition of Plk1 might provide an attractive therapeutic strategy for cancer treatment.p73 is one of newly identified p53 tumor suppressor gene family members (p53, p73, and p63) that encodes a nuclear transcription factor (1-3). Like the other p53 family members, p73 encodes multiple isoforms, including TA (transactivating), with distinct COOH-terminal extensions arising from alternative splicing events and ⌬N (nontransactivating) variants generated by alternative promoter usage (2-4). ⌬Np73 has an oncogenic potential (5) and displays a dominant-negative behavior toward TAp73 as well as wild-type p53 (6). TAp73 transactivates overlapping set of p53-target genes implicated in the induction of cell cycle arrest and/or apoptosis, and plays an important role in the regulation of DNA damage response, which is closely linked to its DNA binding activity. The initial studies demonstrated that TAp73 does not induce enough to accumulate in response to DNA damage arising from UV exposure or actinomycin D treatment (1); however, it has been shown that, in response to certain subset of DNA-damaging agents, TAp73 accumulates in the cell nucleus and exerts its pro-apoptotic function (7).Accumulating evidence suggests that TAp73 is regulated by post-translational modifications such as phosphorylation and acetylation. For example, TAp73 is stabilized in response to DNA damage such as cisplatin (CDDP) 2 treatment or e...

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“…Results from these experiments revealed that only the activities of the β1/β1i and β5/β5i subunits of the proteasome were inhibited by bortezomib [37,38]. The cancer drug, paclitaxel, used to treat breast and ovarian cancers, has also been tested for its efficacy in a cell-based assay using a wortmannin-containing ABP that targets polo-like kinase 1 (PLK1), an enzyme with highly elevated activity in the M phase of the cell cycle [39]. Since paclitaxel arrests the cell cycle at M phase, the elevated activity levels of PLK1 in Jurkat cells treated with this drug were monitored using the wortmannin-containing ABP [39].…”

Section: Enzyme Inhibitor Discovery and Verificationmentioning

confidence: 99%

“…The cancer drug, paclitaxel, used to treat breast and ovarian cancers, has also been tested for its efficacy in a cell-based assay using a wortmannin-containing ABP that targets polo-like kinase 1 (PLK1), an enzyme with highly elevated activity in the M phase of the cell cycle [39]. Since paclitaxel arrests the cell cycle at M phase, the elevated activity levels of PLK1 in Jurkat cells treated with this drug were monitored using the wortmannin-containing ABP [39]. This study demonstrated that PLK1 activity could potentially serve as a biomarker for paclitaxel efficacy in cancer treatment.…”

Section: Enzyme Inhibitor Discovery and Verificationmentioning

confidence: 99%

Application of activity-based probes to the study of enzymes involved in cancer progression

Paulick

Bogyo

2008

Current Opinion in Genetics & Development

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SummaryMany tumor cells have elevated levels of hydrolytic and proteolytic enzymes, presumably to aid in key processes such as angiogenesis, cancer cell invasion, and metastasis. Since the activities of these enzymes are often regulated by post-translational mechanisms, their functional roles in cancer progression are difficult to study using traditional genomic and proteomic methods. Thus, methods that allow for the direct monitoring of enzyme activity in a physiologically relevant environment are required to better understand the roles of specific players in the complex process of tumorigenesis. This review highlights advances in the field of activity-based proteomics, which uses small molecules known as activity-based probes (ABPs) that covalently bind to the catalytic site of target enzymes. We discuss the application of ABPs to cancer biology, specifically to the discovery of tumor biomarkers, the screening of enzyme inhibitors, and the imaging of enzymes implicated in cancer.

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Wortmannin, a Widely Used Phosphoinositide 3-Kinase Inhibitor, also Potently Inhibits Mammalian Polo-like Kinase

Cited by 189 publications

References 51 publications

Rapamycin and mTOR kinase inhibitors

Rapamycin and mTOR kinase inhibitors

Inhibitory Role of Plk1 in the Regulation of p73-dependent Apoptosis through Physical Interaction and Phosphorylation

Application of activity-based probes to the study of enzymes involved in cancer progression

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