S. pombe Aurora Kinase/Survivin Is Required for Chromosome Condensation and the Spindle Checkpoint Attachment Response (original) (raw)
Related papers
Cell, 2000
mitosis and meiosis. Considerable progress has been made in the identification of protein complexes that are required for this event. For example, the condensin complex, which includes the SMC proteins, is thought to facilitate chromosome condensation by imposing topodependent manner (reviewed in Hirano, 2000). However, 1 Department of Biochemistry the molecular mechanisms that link the in vitro activities and Molecular Genetics of the condensin complex with those of other chromo-2 Department of Microbiology somal components in vivo remain to be determined. 3 Department of Chemistry Phosphorylation of histone H3 and linker histone H1 University of Virginia has long been shown to correlate with chromosome Charlottesville, Virginia 22908 condensation during mitosis (Bradbury et al., 1973; Gur-4 Department of Molecular Biology ley et al., 1974). However, since H1 phosphorylation, UT Southwestern Medical Center or even H1 itself, is dispensable for mitotic or meiotic Dallas, Texas 75390 chromosome condensation (Ohsumi et al., 1993; Shen 5 Department of Biochemistry and Molecular Biology et al., 1995), the role for H1 phosphorylation may not be Louisiana State University Medical Center essential for these processes. In contrast, phosphoryla-Shreveport, Louisiana 71130 tion within the amino-terminal domain of H3 is now well 6 Department of Radiation and Cellular Oncology established to be important during mitosis and meiosis in a wide range of organisms (Hendzel et al., 1997; Wei 7 Department of Molecular Genetics et al., 1998). Genetic studies in Tetrahymena show that and Cell Biology a point mutation of serine 10 (Ser-10) in H3 (S10A) leads University of Chicago to abnormal chromosome segregation and extensive Chicago, Illinois 60637 chromosome loss during mitosis and meiosis in micronuclei (Wei et al., 1999). Moreover, inhibiting H3 phosphorylation, presumably by blocking the H3 mitotic Summary kinase activity, prevents the initiation of chromosome condensation and entry into mitosis (Van Hooser et al., 1998; de La Barre et al., 2000)
The EMBO Journal, 2001
We have studied the role of individual histone N-termini and the phosphorylation of histone H3 in chromosome condensation. Nucleosomes, reconstituted with histone octamers containing different combinations of recombinant full-length and tailless histones, were used as competitors for chromosome assembly in Xenopus egg extracts. Nucleosomes reconstituted with intact octamers inhibited chromosome condensation as ef®ciently as the native ones, while tailless nucleosomes were unable to affect this process. Importantly, the addition to the extract of particles containing only intact histone H2B strongly interfered with chromosome formation while such an effect was not observed with particles lacking the N-terminal tail of H2B. This demonstrates that the inhibition effect observed in the presence of competitor nucleosomes is mainly due to the N-terminus of this histone, which, therefore, is essential for chromosome condensation. Nucleosomes in which all histones but H3 were tailless did not impede chromosome formation. In addition, when competitor nucleosome particles were reconstituted with full-length H2A, H2B and H4 and histone H3 mutated at the phosphorylable serine 10 or serine 28, their inhibiting ef®ciency was identical to that of the native particles. Hence, the tail of H3, whether intact or phosphorylated, is not important for chromosome condensation. A novel hypothesis, termed`the ready production label' was suggested to explain the role of histone H3 phosphorylation during cell division.
F1000 - Post-publication peer review of the biomedical literature, 2010
Aurora-B is a component of the Chromosomal Passenger Complex (CPC) required for correct spindle-kinetochore attachments during chromosome segregation and for cytokinesis. The chromatin factors that recruit the CPC to centromeres are unknown, however. Here we show that phosphorylation of Histone-H3 Thr-3 (H3T3ph) by Haspin is necessary for CPC accumulation at centromeres, and that the CPC subunit Survivin binds directly to H3T3ph. A non-binding Survivin-D70A/D71A mutant does not support centromeric CPC concentration and both Haspin depletion and Survivin-D70A/D71A mutation diminish centromere localization of MCAK and mitotic checkpoint signaling in taxol. Survivin-D70A/D71A mutation and microinjection of H3T3ph-specific antibody both compromise centromeric Aurora-B functions but do not prevent cytokinesis. Therefore, H3T3ph generated by Haspin positions the CPC at centromeres to regulate selected targets of Aurora-B during mitosis. The CPC (containing Aurora-B, INCENP, Survivin and Borealin) is found on chromosome arms in prophase, concentrates at inner centromeres during prometaphase, and transfers to the central spindle at anaphase (1). Aurora-B phosphorylates several substrates at these locations, including Histone-H3 Ser-10 (H3S10ph) on chromosome arms, Mitotic Centromere-Associated Kinesin (MCAK) at inner centromeres, Centromere Protein-A Ser-7 (CENP-AS7ph) at outer centromeres and the KNL1/Mis12 complex/Ndc80 network at kinetochores (1-6). Current models suggest that centromeric Aurora-B responds to lack of tension across sister kinetochores that are incorrectly attached to the spindle. Bipolar kinetochore attachment forces may pull kinetochore substrates away from inner centromeric Aurora-B, leading to substrate dephosphorylation, selective stabilization of correct microtubule attachments, and eventually to satisfaction of the spindle checkpoint (Fig. S1) (7). Despite its central importance, it is not understood how Aurora-B accumulates at centromeres (8-10). Immunofluorescence microscopy of mitotic cells shows that H3T3ph and Aurora-B localize similarly at inner centromeres (Fig. 1A, Fig. S2). We therefore tested whether Haspin, which is responsible for generating H3T3ph in mitosis (11,12), is required for CPC localization. Upon Haspin RNAi in HeLa cells, a marked reduction in Aurora-B at centromeres (>5-fold)
Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature, 2010
Aurora-B is a component of the Chromosomal Passenger Complex (CPC) required for correct spindle-kinetochore attachments during chromosome segregation and for cytokinesis. The chromatin factors that recruit the CPC to centromeres are unknown, however. Here we show that phosphorylation of Histone-H3 Thr-3 (H3T3ph) by Haspin is necessary for CPC accumulation at centromeres, and that the CPC subunit Survivin binds directly to H3T3ph. A non-binding Survivin-D70A/D71A mutant does not support centromeric CPC concentration and both Haspin depletion and Survivin-D70A/D71A mutation diminish centromere localization of MCAK and mitotic checkpoint signaling in taxol. Survivin-D70A/D71A mutation and microinjection of H3T3ph-specific antibody both compromise centromeric Aurora-B functions but do not prevent cytokinesis. Therefore, H3T3ph generated by Haspin positions the CPC at centromeres to regulate selected targets of Aurora-B during mitosis. The CPC (containing Aurora-B, INCENP, Survivin and Borealin) is found on chromosome arms in prophase, concentrates at inner centromeres during prometaphase, and transfers to the central spindle at anaphase (1). Aurora-B phosphorylates several substrates at these locations, including Histone-H3 Ser-10 (H3S10ph) on chromosome arms, Mitotic Centromere-Associated Kinesin (MCAK) at inner centromeres, Centromere Protein-A Ser-7 (CENP-AS7ph) at outer centromeres and the KNL1/Mis12 complex/Ndc80 network at kinetochores (1-6). Current models suggest that centromeric Aurora-B responds to lack of tension across sister kinetochores that are incorrectly attached to the spindle. Bipolar kinetochore attachment forces may pull kinetochore substrates away from inner centromeric Aurora-B, leading to substrate dephosphorylation, selective stabilization of correct microtubule attachments, and eventually to satisfaction of the spindle checkpoint (Fig. S1) (7). Despite its central importance, it is not understood how Aurora-B accumulates at centromeres (8-10). Immunofluorescence microscopy of mitotic cells shows that H3T3ph and Aurora-B localize similarly at inner centromeres (Fig. 1A, Fig. S2). We therefore tested whether Haspin, which is responsible for generating H3T3ph in mitosis (11,12), is required for CPC localization. Upon Haspin RNAi in HeLa cells, a marked reduction in Aurora-B at centromeres (>5-fold)
The Journal of Cell Biology, 2001
Aurora/Ipl1-related kinases are a conserved family of enzymes that have multiple functions during mitotic progression. Although it has been possible to use conventional genetic analysis to dissect the function of aurora , the founding family member in Drosophila (Glover, D.M., M.H. Leibowitz, D.A. McLean, and H. Parry. 1995. Cell . 81:95-105), the lack of mutations in a second aurora-like kinase gene, aurora B , precluded this approach. We now show that depleting Aurora B kinase using double-stranded RNA interference in cultured Drosophila cells results in polyploidy. aurora B encodes a passenger protein that associates first with condensing chromatin, concentrates at centromeres, and then relocates onto the central spindle at anaphase. Cells depleted of the Aurora B kinase show only partial chromosome condensation at mitosis. This is associated with a reduction in levels of the serine 10 phosphorylated form of histone H3 and a failure to recruit the Barren condensin protein onto chromosomes. These defects are associated with abnormal segregation resulting from lagging chromatids and extensive chromatin bridging at anaphase, similar to the phenotype of barren mutants (Bhat, M.A., A.V. . Cell . 87:1103-1114. The majority of treated cells also fail to undertake cytokinesis and show a reduced density of microtubules in the central region of the spindle. This is accompanied by a failure to correctly localize the Pavarotti kinesinlike protein, essential for this process. We discuss these conserved functions of Aurora B kinase in chromosome transmission and cytokinesis.
Molecular Cell, 2000
with the help of motor proteins . Spindle checkpoints ensure that only one member of each pair of sister chromatids moves to a particular pole by preventing the activation of the APC until all chromosomes are aligned at the metaphase plate and Building 68 Room 425 are connected to microtubules at their kinetochores (re-Massachusetts Institute of Technology viewed by Amon, 1999). Between the segregating sister Cambridge, Massachusetts 02139 chromatids in anaphase, a tubulin-containing structure † The Walter and Eliza Hall Institute of Medical Research called the spindle midzone forms. Proteins that localize Post Office Royal Melbourne Hospital to the spindle midzone may affect cytokinesis by speci-Victoria 3050 fying the site of formation of the cytokinetic furrow or Australia by promoting furrow ingression or resolution (reviewed by Glotzer, 1997). Following cytokinesis, the cytokinetic remnant, known as the midbody in mammals, remains Summary at the site where the cytokinetic furrow divides the cell at the spindle midzone. Phosphorylation and dephosphorylation events are Baculoviral IAP repeat proteins (BIRPs) may affect cell critical for the cytoskeletal and chromosomal changes death, cell division, and tumorigenesis. The C. elegans that occur during mitosis. The dissolution of the nuclear BIRP BIR-1 was localized to chromosomes and to the membrane, condensation of chromosomes, duplication spindle midzone. Embryos and fertilized oocytes lackof the spindle poles, and cytokinesis have been correing BIR-1 had defects in chromosome behavior, spinlated with the phosphorylation of putative effectors, indle midzone formation, and cytokinesis. We observed cluding lamins, histones, Cut12p, and myosin II, respecindistinguishable defects in fertilized oocytes and emtively (Satterwhite et al., 1992; Bridge et al., 1998; Collas, bryos lacking the Aurora-like kinase AIR-2. AIR-2 was 1999; Wei et al., 1999). The Cdc2, Polo, and Aurora not present on chromosomes in the absence of BIR-1.
Journal of Cell Science, 2001
Metazoans contain three aurora-related kinases. Aurora A is required for spindle formation while aurora B is required for chromosome condensation and cytokinesis. Less is known about the function of aurora C. S. pombe contains a single aurora-related kinase, Ark1. Although Ark1 protein levels remained constant as cells progressed through the mitotic cell cycle, its distribution altered during mitosis and meiosis. Throughout G2 Ark1 was concentrated in one to three nuclear foci that were not associated with the spindle pole body/centromere complex. Following commitment to mitosis Ark1 associated with chromatin and was particularly concentrated at several sites including kinetochores/centromeres. Kinetochore/centromere association diminished during anaphase A, after which it was distributed along the spindle. The protein became restricted to a small central zone that transiently enlarged as the spindle extended. As in many other systems mitotic fission yeast cells exhibit a much great...
Global Role for Chromatin Remodeling Enzymes in Mitotic Gene Expression
Cell, 2000
Swi5p also binds to the HO Worcester, Massachusetts 01605 promoter during late mitosis, although HO is not expressed until late G1 (Cosma et al., 1999). Ace2p is a zinc finger protein that is highly homolo-Summary gous to Swi5p and that regulates transcription of many of the same genes. These two proteins share 83% iden-Regulation of eukaryotic gene expression requires tity between their zinc finger domains, and they recog-ATP-dependent chromatin remodeling enzymes, such nize similar binding sites in vitro (Dohrmann et al., 1992). as SWI/SNF, and histone acetyltransferases, such as Ace2p is also subject to similar cell cycle regulation Gcn5p. Here we show that SWI/SNF remodeling conas Swi5p, both in terms of expression during G2 and trols recruitment of Gcn5p HAT activity to many genes regulated nuclear localization during late anaphase in late mitosis and that these chromatin remodeling (Dohrmann et al., 1992; O'Conallain et al., 1999). Despite these similarities, Swi5p and Ace2p play distinct roles enzymes play a role in regulating mitotic exit. In conin mitotic gene expression. Whereas Swi5p activates trast, interphase expression of GAL1, HIS3, PHO5, and target genes such as SIC1 and PCL9 immediately after PHO8 is accompanied by SWI/SNF-independent reentering the nucleus in late anaphase, Ace2p is inactive cruitment of Gcn5p HAT activity. Surprisingly, prearduring mitosis and can activate expression of these resting cells in late mitosis imposes a requirement for same genes only during early G1 (Aerne et al., 1998; SWI/SNF in recruiting Gcn5p HAT activity to the GAL1 McBride et al., 1999).
An acetylated form of histone H2A.Z regulates chromosome architecture in Schizosaccharomyces pombe
Nature Structural & Molecular Biology, 2009
Histone variant H2A.Z has a conserved role in genome stability, although it remains unclear how this is mediated. Here we demonstrate that the fission yeast Swr1 ATPase inserts H2A.Z (Pht1) into chromatin and Kat5 acetyltransferase (Mst1) acetylates it. Deletion or an unacetylatable mutation of Pht1 leads to genome instability, primarily caused by chromosome entanglement and breakage at anaphase. This leads to the loss of telomere-proximal markers, though telomere protection and repeat length are unaffected by the absence of Pht1. Strikingly, the chromosome entanglement in pht1∆ anaphase cells can be rescued by forcing chromosome condensation before anaphase onset. We show that the condensin complex, required for the maintenance of anaphase chromosome condensation, prematurely dissociates from chromatin in the absence of Pht1. This and other findings suggest an important role for H2A.Z in the architecture of anaphase chromosomes.