XB-ART-9884
Mol Biol Cell
2000 Dec 01;1112:4173-87. doi: 10.1091/mbc.11.12.4173.
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Identification of ribonucleotide reductase protein R1 as an activator of microtubule nucleation in Xenopus egg mitotic extracts.
Takada S
,
Shibata T
,
Hiraoka Y
,
Masuda H
.
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Microtubule nucleation on the centrosome and the fungal equivalent, the spindle pole body (SPB), is activated at the onset of mitosis. We previously reported that mitotic extracts prepared from Xenopus unfertilized eggs convert the interphase SPB of fission yeast into a competent state for microtubule nucleation. In this study, we have purified an 85-kDa SPB activator from the extracts and identified it as the ribonucleotide reductase large subunit R1. We further confirmed that recombinant mouse R1 protein was also effective for SPB activation. On the other hand, another essential subunit of ribonucleotide reductase, R2 protein, was not required for SPB activation. SPB activation by R1 protein was suppressed in the presence of anti-R1 antibodies or a partial oligopeptide of R1; the oligopeptide also inhibited aster formation on Xenopus sperm centrosomes. In accordance, R1 was detected in animal centrosomes by immunofluorescence and immunoblotting with anti-R1 antibodies. In addition, recombinant mouse R1 protein bound to gamma- and alpha/beta-tubulin in vitro. These results suggest that R1 is a bifunctional protein that acts on both ribonucleotide reduction and centrosome/SPB activation.
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References [+] :
Bailly,
p34cdc2 is located in both nucleus and cytoplasm; part is centrosomally associated at G2/M and enters vesicles at anaphase.
1989, Pubmed
Bailly, p34cdc2 is located in both nucleus and cytoplasm; part is centrosomally associated at G2/M and enters vesicles at anaphase. 1989, Pubmed
Bailly, Cytoplasmic accumulation of cyclin B1 in human cells: association with a detergent-resistant compartment and with the centrosome. 1992, Pubmed
Bornens, Structural and chemical characterization of isolated centrosomes. 1987, Pubmed , Xenbase
Brinkley, Supernumerary centrosomes and cancer: Boveri's hypothesis resurrected. 1998, Pubmed
Brown, Both viral (adenovirus E1B) and cellular (hsp 70, p53) components interact with centrosomes. 1994, Pubmed
Brown, Role of effector binding in allosteric control of ribonucleoside diphosphate reductase. 1969, Pubmed
Buendia, Regulation of the microtubule nucleating activity of centrosomes in Xenopus egg extracts: role of cyclin A-associated protein kinase. 1992, Pubmed , Xenbase
Caras, Cloned mouse ribonucleotide reductase subunit M1 cDNA reveals amino acid sequence homology with Escherichia coli and herpesvirus ribonucleotide reductases. 1985, Pubmed
Chabes, Yeast Sml1, a protein inhibitor of ribonucleotide reductase. 1999, Pubmed
Davis, Purification, characterization, and localization of subunit interaction area of recombinant mouse ribonucleotide reductase R1 subunit. 1994, Pubmed
Dictenberg, Pericentrin and gamma-tubulin form a protein complex and are organized into a novel lattice at the centrosome. 1998, Pubmed , Xenbase
Ding, The spindle pole body of Schizosaccharomyces pombe enters and leaves the nuclear envelope as the cell cycle proceeds. 1997, Pubmed
Doxsey, Pericentrin, a highly conserved centrosome protein involved in microtubule organization. 1994, Pubmed , Xenbase
Elledge, DNA damage and cell cycle regulation of ribonucleotide reductase. 1993, Pubmed
Engström, Cell cycle-dependent expression of mammalian ribonucleotide reductase. Differential regulation of the two subunits. 1985, Pubmed
Engström, Localization of ribonucleotide reductase in mammalian cells. 1984, Pubmed
Erickson, Protofilaments and rings, two conformations of the tubulin family conserved from bacterial FtsZ to alpha/beta and gamma tubulin. 1996, Pubmed
Eriksson, Binding of allosteric effectors to ribonucleotide reductase protein R1: reduction of active-site cysteines promotes substrate binding. 1997, Pubmed
Eriksson, Cell cycle-dependent regulation of mammalian ribonucleotide reductase. The S phase-correlated increase in subunit M2 is regulated by de novo protein synthesis. 1984, Pubmed
Fava, Human 76p: A new member of the gamma-tubulin-associated protein family. 1999, Pubmed , Xenbase
Félix, Centrosome assembly in vitro: role of gamma-tubulin recruitment in Xenopus sperm aster formation. 1994, Pubmed , Xenbase
Fernandez Sarabia, The cell cycle genes cdc22+ and suc22+ of the fission yeast Schizosaccharomyces pombe encode the large and small subunits of ribonucleotide reductase. 1993, Pubmed
Gallant, Cyclin B2 undergoes cell cycle-dependent nuclear translocation and, when expressed as a non-destructible mutant, causes mitotic arrest in HeLa cells. 1992, Pubmed , Xenbase
Geissler, The spindle pole body component Spc98p interacts with the gamma-tubulin-like Tub4p of Saccharomyces cerevisiae at the sites of microtubule attachment. 1996, Pubmed
Hagan, The use of cell division cycle mutants to investigate the control of microtubule distribution in the fission yeast Schizosaccharomyces pombe. 1988, Pubmed
Horio, Human gamma-tubulin functions in fission yeast. 1994, Pubmed
Horio, The fission yeast gamma-tubulin is essential for mitosis and is localized at microtubule organizing centers. 1991, Pubmed
Huang, The DNA replication and damage checkpoint pathways induce transcription by inhibition of the Crt1 repressor. 1998, Pubmed
Huang, Identification of RNR4, encoding a second essential small subunit of ribonucleotide reductase in Saccharomyces cerevisiae. 1997, Pubmed
Hyman, Preparation of modified tubulins. 1991, Pubmed
Ingemarson, A kinetic study on the influence of nucleoside triphosphate effectors on subunit interaction in mouse ribonucleotide reductase. 1996, Pubmed
Joshi, Gamma-tubulin is a centrosomal protein required for cell cycle-dependent microtubule nucleation. 1992, Pubmed
Kalt, Molecular components of the centrosome. 1993, Pubmed
Kellogg, The centrosome and cellular organization. 1994, Pubmed
Khodjakov, The sudden recruitment of gamma-tubulin to the centrosome at the onset of mitosis and its dynamic exchange throughout the cell cycle, do not require microtubules. 1999, Pubmed
Knop, The spindle pole body component Spc97p interacts with the gamma-tubulin of Saccharomyces cerevisiae and functions in microtubule organization and spindle pole body duplication. 1997, Pubmed
Kuriyama, Microtubule-nucleating activity of centrosomes in Chinese hamster ovary cells is independent of the centriole cycle but coupled to the mitotic cycle. 1981, Pubmed
Maldonado-Codina, Cyclins A and B associate with chromatin and the polar regions of spindles, respectively, and do not undergo complete degradation at anaphase in syncytial Drosophila embryos. 1992, Pubmed
Marschall, Analysis of Tub4p, a yeast gamma-tubulin-like protein: implications for microtubule-organizing center function. 1996, Pubmed
Martin, Xgrip109: a gamma tubulin-associated protein with an essential role in gamma tubulin ring complex (gammaTuRC) assembly and centrosome function. 1998, Pubmed , Xenbase
Masuda, Role of gamma-tubulin in mitosis-specific microtubule nucleation from the Schizosaccharomyces pombe spindle pole body. 1996, Pubmed , Xenbase
Masuda, In vitro microtubule-nucleating activity of spindle pole bodies in fission yeast Schizosaccharomyces pombe: cell cycle-dependent activation in xenopus cell-free extracts. 1992, Pubmed , Xenbase
Masuda, In vitro reactivation of spindle elongation in fission yeast nuc2 mutant cells. 1990, Pubmed
Moritz, Recruitment of the gamma-tubulin ring complex to Drosophila salt-stripped centrosome scaffolds. 1998, Pubmed , Xenbase
Moritz, Microtubule nucleation by gamma-tubulin-containing rings in the centrosome. 1995, Pubmed
Moudjou, gamma-Tubulin in mammalian cells: the centrosomal and the cytosolic forms. 1996, Pubmed
Moynihan, Liz1p, a novel fission yeast membrane protein, is required for normal cell division when ribonucleotide reductase is inhibited. 1999, Pubmed
Murphy, The mammalian gamma-tubulin complex contains homologues of the yeast spindle pole body components spc97p and spc98p. 1998, Pubmed , Xenbase
Murray, Cell cycle extracts. 1991, Pubmed
Nakamura, When overexpressed, a novel centrosomal protein, RanBPM, causes ectopic microtubule nucleation similar to gamma-tubulin. 1998, Pubmed
Nasmyth, Cell division cycle mutants altered in DNA replication and mitosis in the fission yeast Schizosaccharomyces pombe. 1981, Pubmed
Oakley, Gamma-tubulin is a component of the spindle pole body that is essential for microtubule function in Aspergillus nidulans. 1990, Pubmed
Oakley, Gamma-tubulin: the microtubule organizer? 1992, Pubmed
Ohta, Microtubule nucleating activity of centrosomes in cell-free extracts from Xenopus eggs: involvement of phosphorylation and accumulation of pericentriolar material. 1993, Pubmed , Xenbase
Parker, Human M1 subunit of ribonucleotide reductase: cDNA sequence and expression in stimulated lymphocytes. 1991, Pubmed
Pennisi, Nuclear transport protein does double duty in mitosis. 1999, Pubmed , Xenbase
Pereira, Centrosome-microtubule nucleation. 1997, Pubmed , Xenbase
Pereira, Spc98p directs the yeast gamma-tubulin complex into the nucleus and is subject to cell cycle-dependent phosphorylation on the nuclear side of the spindle pole body. 1998, Pubmed
Reichard, From RNA to DNA, why so many ribonucleotide reductases? 1993, Pubmed
Reichard, Interactions between deoxyribonucleotide and DNA synthesis. 1988, Pubmed
Reinsch, Mechanisms of nuclear positioning. 1998, Pubmed
Sibon, DNA-replication/DNA-damage-dependent centrosome inactivation in Drosophila embryos. 2000, Pubmed
Snyder, Initiation and growth of microtubules from mitotic centers in lysed mammalian cells. 1975, Pubmed
Sobel, A highly divergent gamma-tubulin gene is essential for cell growth and proper microtubule organization in Saccharomyces cerevisiae. 1995, Pubmed
Spang, gamma-Tubulin-like Tub4p of Saccharomyces cerevisiae is associated with the spindle pole body substructures that organize microtubules and is required for mitotic spindle formation. 1996, Pubmed , Xenbase
Stearns, In vitro reconstitution of centrosome assembly and function: the central role of gamma-tubulin. 1994, Pubmed , Xenbase
Sunkel, Gamma-tubulin is required for the structure and function of the microtubule organizing centre in Drosophila neuroblasts. 1995, Pubmed
Tanaka, A ribonucleotide reductase gene involved in a p53-dependent cell-cycle checkpoint for DNA damage. 2000, Pubmed
Tassin, Characterization of the human homologue of the yeast spc98p and its association with gamma-tubulin. 1998, Pubmed , Xenbase
Thelander, Ribonucleotide reductase from calf thymus. Separation of the enzyme into two nonidentical subunits, proteins M1 and M2. 1980, Pubmed
Thelander, Reduction of ribonucleotides. 1979, Pubmed
Thelander, Subunit M2 of mammalian ribonucleotide reductase. Characterization of a homogeneous protein isolated from M2-overproducing mouse cells. 1985, Pubmed
Tugendreich, CDC27Hs colocalizes with CDC16Hs to the centrosome and mitotic spindle and is essential for the metaphase to anaphase transition. 1995, Pubmed
Uhlin, Structure of ribonucleotide reductase protein R1. 1994, Pubmed
Vardy, The fission yeast gamma-tubulin complex is required in G(1) phase and is a component of the spindle assembly checkpoint. 2000, Pubmed
Vernos, Motors involved in spindle assembly and chromosome segregation. 1996, Pubmed
Vogel, Centrosomes isolated from Spisula solidissima oocytes contain rings and an unusual stoichiometric ratio of alpha/beta tubulin. 1997, Pubmed
Wang, Rnr4p, a novel ribonucleotide reductase small-subunit protein. 1997, Pubmed
Waters, Pathways of spindle assembly. 1997, Pubmed
Wigley, Dynamic association of proteasomal machinery with the centrosome. 1999, Pubmed
Zhao, A suppressor of two essential checkpoint genes identifies a novel protein that negatively affects dNTP pools. 1998, Pubmed
Zheng, Nucleation of microtubule assembly by a gamma-tubulin-containing ring complex. 1995, Pubmed , Xenbase