Characterization of apoptosis-induced Mcm3 and Cdc6 cleavage reveals a proapoptotic effect for one Mcm3 fragment

Schories, Barbara; Engel, Katrin; Dörken, Bernd; Gossen, Manfred; Bommert, Kurt

doi:10.1038/sj.cdd.4401411

Cited by 17 publications

(12 citation statements)

References 9 publications

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“…In budding yeast these apoptotic triggers include extensive DNA damage (Blanchard et al, 2002;Qi et al, 2003;Weinberger et al, 2005) and defects in the replication-initiation proteins Cdc6p (Blanchard et al, 2002) and Orc2p (Weinberger et al, 2005). The same triggers, DNA damage (Norbury and Zhivotovsky, 2004) and defects in replication-initiation proteins (Blanchard et al, 2002;Dodson et al, 2004;Feng et al, 2003;Kim et al, 2003;Pelizon et al, 2002;Schories et al, 2004;Shreeram et al, 2002;Yim et al, 2003), can also induce apoptosis in mammalian cells.…”

Section: Introductionmentioning

confidence: 99%

Production of reactive oxygen species in response to replication stress and inappropriate mitosis in fission yeast

Marchetti

Weinberger

Murakami

et al. 2006

Journal of Cell Science

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Previous studies have indicated that replication stress can trigger apoptosis-like cell death, accompanied (where tested) by production of reactive oxygen species (ROS), in mammalian cells and budding yeast (Saccharomyces cerevisiae). In mammalian cells, inappropriate entry into mitosis also leads to cell death. Here, we report similar responses in fission yeast (Schizosaccharomyces pombe). We used ROS- and death-specific fluorescent stains to measure the effects of mutations in replication initiation and checkpoint genes in fission yeast on the frequencies of ROS production and cell death. We found that certain mutant alleles of each of the four tested replication initiation genes caused elevated ROS and cell death. Where tested, these effects were not enhanced by checkpoint-gene mutations. Instead, when cells competent for replication but defective in both the replication and damage checkpoints were treated with hydroxyurea, which slows replication fork movement, the frequencies of ROS production and cell death were greatly increased. This was a consequence of elevated CDK activity, which permitted inappropriate entry into mitosis. Thus, studies in fission yeast are likely to prove helpful in understanding the pathways that lead from replication stress and inappropriate mitosis to cell death in mammalian cells.

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Section: Introductionmentioning

confidence: 99%

Production of reactive oxygen species in response to replication stress and inappropriate mitosis in fission yeast

Marchetti

Weinberger

Murakami

et al. 2006

Journal of Cell Science

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“…One of the highest scoring candidates for S9 identified by mass spectrometry was MCM3, a component of the replication licensing complex that undergoes caspase-mediated cleavage during apoptosis (25,26). Addition of DK1 to FLAG-tagged MCM3 led to significant…”

Section: Resultsmentioning

confidence: 99%

“…FLAG-tagged L5 was generated by PCR cloning of the rat L5 cDNA (a kind gift from O. Meyuhas, Hebrew University, Jerusalem, Israel) into pcDNA3. FLAG-tagged human MCM3 in pcDNA3 was a kind gift from Dr. M. Gossen (Max Delbruck Center for Molecular Medicine, Berlin, Germany) (25). Ser 160 was mutated to either Ala or Asp by PCR-based site-directed mutagenesis using the QuikChange mutagenesis kit (Stratagene) according to the recommended protocol.…”

Section: Methodsmentioning

confidence: 99%

A High Throughput Proteomics Screen Identifies Novel Substrates of Death-associated Protein Kinase

Bialik

Berissi

Kimchi

2008

Molecular & Cellular Proteomics

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Death-associated protein kinase (DAPk) is a Ser/Thr kinase whose activity is necessary for different cell death phenotypes. Although its contribution to cell death is well established, only a handful of direct substrates have been identified; these do not fully account for the multiple cellular effects of DAPk. To identify such substrates on a large scale, we developed an in vitro, unbiased, proteomics-based assay to search for novel DAPk substrates. Biochemical fractionation and mass spectrometric analysis were used to purify and identify several potential substrates from HeLa cell lysate. Here we report the identification of two such candidate substrates, the ribosomal protein L5 and MCM3, a replication licensing factor. Although L5 proved to be a weak substrate, MCM3 was efficiently and specifically phosphorylated by DAPk on a unique site, Ser 160 . Significantly DAPk phosphorylated this site in vivo upon overexpression in 293T cells. Activation of endogenous DAPk by increasing intracellular Ca 2؉ also led to increased phosphorylation of MCM3. Importantly short hairpin RNAmediated knockdown of endogenous DAPk blocked both basal phosphorylation and Ca 2؉ -induced phosphorylation, indicating that DAPk is both necessary and sufficient for MCM3 Ser 160 phosphorylation in vivo. Identification of MCM3 as an in vivo DAPk substrate indicates the usefulness of this approach for identification of physiologically relevant substrates that may shed light on novel functions of the kinase.

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“…MCM3 has been implicated in the regulation of the eukaryotic MCM complex [4], and has been shown to be specifically phosphorylated by ATM/ATR kinases [10]. An apparent requirement for the specific proteolysis of eukaryotic MCM3 before apoptosis can be induced has also been reported [48,49]. Thus the notion of a specialized Mcm as a nexus for a modulatory or checkpoint decision is not without precedent.…”

Section: Discussionmentioning

confidence: 99%

Non-essential MCM-related proteins mediate a response to DNA damage in the archaeon Methanococcus maripaludis

Walters

Chong

2017

Microbiology

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The single minichromosome maintenance (MCM) protein found in most archaea has been widely studied as a simplified model for the MCM complex that forms the catalytic core of the eukaryotic replicative helicase. Organisms of the order Methanococcales are unusual in possessing multiple MCM homologues. The Methanococcus maripaludis S2 genome encodes four MCM homologues, McmA-McmD. DNA helicase assays reveal that the unwinding activity of the three MCM-like proteins is highly variable despite sequence similarities and suggests additional motifs that influence MCM function are yet to be identified. While the gene encoding McmA could not be deleted, strains harbouring individual deletions of genes encoding each of the other MCMs display phenotypes consistent with these proteins modulating DNA damage responses. M. maripaludis S2 is the first archaeon in which MCM proteins have been shown to influence the DNA damage response.

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Characterization of apoptosis-induced Mcm3 and Cdc6 cleavage reveals a proapoptotic effect for one Mcm3 fragment

Cited by 17 publications

References 9 publications

Production of reactive oxygen species in response to replication stress and inappropriate mitosis in fission yeast

Production of reactive oxygen species in response to replication stress and inappropriate mitosis in fission yeast

A High Throughput Proteomics Screen Identifies Novel Substrates of Death-associated Protein Kinase

Non-essential MCM-related proteins mediate a response to DNA damage in the archaeon Methanococcus maripaludis

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