Participation of a Fusogenic Protein, Glyceraldehyde-3-phosphate Dehydrogenase, in Nuclear Membrane Assembly

Nakagawa, Toshimasa; Hirano, Yasuhiro; Inomata, Akira; Yokota, Sadaki; Miyachi, Kiyomitsu; Kaneda, Mizuho; Umeda, Masato; Furukawa, Kazuhiro; Omata, Saburo; Horigome, Tsuneyoshi

doi:10.1074/jbc.m210824200

Cited by 60 publications

(40 citation statements)

References 55 publications

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“…These include the transcriptional regulation of histone gene expression (Zheng et al, 2003), the role of GAPDH in nuclear membrane fusion (Nakagawa et al, 2003), in the modulation of telomere structure (Sundararaj et al, 2004), in the recognition of fraudulent nucleotides in DNA arising from cancer chemotherapy (Krynetski et al, 2003) and in triggering apoptosis (Chuang et al, 2005;Hara et al, 2005).…”

Section: Discussionmentioning

confidence: 99%

Glyceraldehyde 3-phosphate dehydrogenase is a SET-binding protein and regulates cyclin B-cdk1 activity

et al. 2006

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We report here that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) interacts in vitro and in vivo with the protein SET. This interaction is performed through the acidic domain of SET located at the carboxy terminal region. On analysing the functional relevance of SET-GAPDH interaction, we observed that GAPDH reverses in a dose-dependent manner, the inhibition of cyclin Bcdk1 activity produced by SET. Similarly to SET, GAPDH associates with cyclin B, suggesting that the regulation of cyclin B-cdk1 activity might be mediated not only by the interaction of GAPDH with SET but also with cyclin B. To analyse the putative role of GAPDH on cell cycle progression, HCT116 cells were transfected with a GAPDH expression vector. Results indicate that overexpression of GAPDH does not affect the timing of DNA replication but induces an increase in the number of mitosis, an advancement of the peak of cyclin B-cdk1 activity and an acceleration of cell cycle progression. All these results suggest that GAPDH might be involved in cell cycle regulation by modulating cyclin B-cdk1 activity.

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Section: Discussionmentioning

confidence: 99%

Glyceraldehyde 3-phosphate dehydrogenase is a SET-binding protein and regulates cyclin B-cdk1 activity

et al. 2006

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“…GAPDH is an enzyme of glycolysis, but is also involved in eukaryotic membrane fusion events, including fusion of endoplasmic reticulum-derived vesicles with the Golgi apparatus (Glaser & Gross, 1995;Nakagawa et al, 2003;Tisdale, 2001;Tisdale et al, 2004). The addition of GAPDH stimulated killing of E. coli by mxOMVs, suggesting that predatory activity was mediated by fusion with the E. coli membrane, as suggested for OMVs of Pseudomonas aeruginosa (Kadurugamuwa & Beveridge, 1996).…”

Section: A G L Evans and Othersmentioning

confidence: 99%

Predatory activity of Myxococcus xanthus outer-membrane vesicles and properties of their hydrolase cargo

et al. 2012

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The deltaproteobacterium Myxococcus xanthus predates upon members of the soil microbial community by secreting digestive factors and lysing prey cells. Like other Gram-negative bacteria, M. xanthus produces outer membrane vesicles (OMVs), and we show here that M. xanthus OMVs are able to kill Escherichia coli cells. The OMVs of M. xanthus were found to contain active proteases, phosphatases, other hydrolases and secondary metabolites. Alkaline phosphatase activity was found to be almost exclusively associated with OMVs, implying that there is active targeting of phosphatases into OMVs, while other OMV components appear to be packaged passively. The kinetic properties of OMV alkaline phosphatase suggest that there may have been evolutionary adaptation of OMV enzymes to a relatively indiscriminate mode of action, consistent with a role in predation. In addition, the observed regulation of production, and fragility of OMV activity, may protect OMV-producing cells from exploitation by M. xanthus cheating genotypes and/or other competitors. Killing of E. coli by M. xanthus OMVs was enhanced by the addition of a fusogenic enzyme (glyceraldehyde-3-phosphate dehydrogenase; GAPDH), which triggers fusion of vesicles with target membranes within eukaryotic cells. This suggests that the mechanism of prey killing involves OMV fusion with the E. coli outer membrane. M. xanthus secretes GAPDH, which could potentially modulate the fusion of co-secreted OMVs with prey organisms in nature, enhancing their predatory activity.

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“…Preparation of Xenopus Egg Cytosol Fractions-Xenopus eggs were collected, dejelled, and then lysed to prepare S-phase and M-phase cytosol fractions as described previously (26,27).…”

Section: Construction Of Gst-fused Emerin Fragment Proteins and Hismentioning

confidence: 99%

Dissociation of Emerin from Barrier-to-autointegration Factor Is Regulated through Mitotic Phosphorylation of Emerin in a Xenopus Egg Cell-free System

Hirano

Segawa

Ouchi

et al. 2005

Journal of Biological Chemistry

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Emerin is the gene product of STA whose mutations cause Emery-Dreifuss muscular dystrophy. It is an inner nuclear membrane protein and phosphorylated in a cell cycle-dependent manner. However, the means of phosphorylation of emerin are poorly understood. We investigated the regulation mechanism for the binding of emerin to chromatin, focusing on its cell cycle-dependent phosphorylation in a Xenopus egg cell-free system. It was shown that emerin dissociates from chromatin depending on mitotic phosphorylation of the former, and this plays a critical role in the dissociation of emerin from barrier-to-autointegration factor (BAF). Then, we analyzed the mitotic phosphorylation sites of emerin. Emerin was strongly phosphorylated in an M-phase Xenopus egg cell-free system, and five phosphorylated sites, Ser 49 , Ser 66 , Thr 67 , Ser 120 , and Ser 175 , were identified on analysis of chymotryptic and tryptic emerin peptides using a phosphopeptide-concentrating system coupled with a Titansphere column, which specifically binds phosphopeptides, and tandem mass spectrometry sequencing. An in vitro binding assay involving an emerin S175A point mutant protein suggested that phosphorylation at Ser 175 regulates the dissociation of emerin from BAF. The nuclear envelope (NE)2 is a highly dynamic structure that disassembles at the onset of mitosis and reassembles on the surface of chromatin during telophase in vertebrates. These changes of NE are crucial for cell cycle progression. The NE consists of an outer nuclear membrane, inner nuclear membrane, nuclear pore complex, and nuclear lamina. The inner nuclear membrane contains integral membrane proteins, i.e. lamin B receptor (LBR), lamina-associated polypeptide-2␤ (LAP2␤), emerin, MAN1, and others, which interact with DNA and/or chromatin, and these proteins are proposed to participate in nuclear membrane targeting to chromatin at an early stage of nuclear assembly (1). The interactions between some of the inner nuclear membrane proteins and chromatin are regulated through phosphorylation of these inner nuclear membrane proteins. The phosphorylation mechanisms for LBR and LAP2␣ and 2␤ are well understood (2-7). LBR directly binds to DNA in vitro and dissociates on phosphorylation by cdc2 kinase and other kinase(s) in a mitotic egg extract (3). LAP2␤ binds to lamin B1 and chromatin, and cell cycle-dependent phosphorylation of LAP2␤ cancels this binding (4). Phosphorylation of these inner nuclear proteins, therefore, is likely to be one of the key mechanisms that control the interactions between the inner nuclear proteins and components of the nuclear lamina as well as chromatin. In this study, we focused on the mitotic phosphorylation of emerin, one of the inner nuclear membrane proteins.Human emerin is a serine-rich protein exhibiting an apparent mass of 34 kDa on SDS-PAGE (8) and is phosphorylated in a cell cycle-dependent manner (9). Emerin belongs to the LEM (LAP2␤, emerin, MAN1) protein family, whose members have approximately a 40-residue domain named the LEM (10). These ...

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Participation of a Fusogenic Protein, Glyceraldehyde-3-phosphate Dehydrogenase, in Nuclear Membrane Assembly

Cited by 60 publications

References 55 publications

Glyceraldehyde 3-phosphate dehydrogenase is a SET-binding protein and regulates cyclin B-cdk1 activity

Glyceraldehyde 3-phosphate dehydrogenase is a SET-binding protein and regulates cyclin B-cdk1 activity

Predatory activity of Myxococcus xanthus outer-membrane vesicles and properties of their hydrolase cargo

Dissociation of Emerin from Barrier-to-autointegration Factor Is Regulated through Mitotic Phosphorylation of Emerin in a Xenopus Egg Cell-free System

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