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1.  Direct kinetochore–spindle pole connections are not required for chromosome segregation 
The Journal of Cell Biology  2014;206(2):231-243.
In the absence of continuous K-fiber attachment between each kinetochore and the spindle pole, one or more additional mechanisms dependent on dynein-mediated kinetochore transport exist to ensure proper chromosome segregation during mitosis.
Segregation of genetic material occurs when chromosomes move to opposite spindle poles during mitosis. This movement depends on K-fibers, specialized microtubule (MT) bundles attached to the chromosomes′ kinetochores. A long-standing assumption is that continuous K-fibers connect every kinetochore to a spindle pole and the force for chromosome movement is produced at the kinetochore and coupled with MT depolymerization. However, we found that chromosomes still maintained their position at the spindle equator during metaphase and segregated properly during anaphase when one of their K-fibers was severed near the kinetochore with a laser microbeam. We also found that, in normal fully assembled spindles, K-fibers of some chromosomes did not extend to the spindle pole. These K-fibers connected to adjacent K-fibers and/or nonkinetochore MTs. Poleward movement of chromosomes with short K-fibers was uncoupled from MT depolymerization at the kinetochore. Instead, these chromosomes moved by dynein-mediated transport of the entire K-fiber/kinetochore assembly. Thus, at least two distinct parallel mechanisms drive chromosome segregation in mammalian cells.
doi:10.1083/jcb.201401090
PMCID: PMC4107786  PMID: 25023516
2.  Circumventing photodamage in live-cell microscopy 
Methods in cell biology  2013;114:10.1016/B978-0-12-407761-4.00023-3.
Fluorescence microscopy has become an essential tool in cell biology. This technique allows researchers to visualize the dynamics of tissue, cells, individual organelles and macromolecular assemblies inside the cell. Unfortunately, fluorescence microscopy is not completely ‘non-invasive’ as the high-intensity excitation light required for excitation of fluorophores is inherently toxic for live cells. Physiological changes induced by excessive illumination can lead to artifacts and abnormal responses. In this chapter we review major factors that contribute to phototoxicity and discuss practical solutions for circumventing photodamage. These solutions include the proper choice of image acquisition parameters, optimization of filter sets, hardware synchronization, and the use of intelligent illumination to avoid unnecessary light exposure.
doi:10.1016/B978-0-12-407761-4.00023-3
PMCID: PMC3843244  PMID: 23931522
fluorescence; live-cell microscopy; phototoxicity; photodamage; GFP
3.  A kinesin-mediated mechanism that couples centrosomes to nuclei 
The M-type kinesin isoform, Kif9, has recently been implicated in maintaining a physical connection between the centrosome and nucleus in Dictyostelium discoideum. However, the mechanism by which Kif9 functions to link these two organelles remains obscure. Here we demonstrate that the Kif9 protein is localized to the nuclear envelope and is concentrated in the region underlying the centrosome point of attachment. Nuclear anchorage appears mediated through a specialized trans-membrane domain located in the carboxyl terminus. Kif9 interacts with microtubules in in vitro binding assays and effects an endwise depolymerization of the polymer. These results suggest a model whereby Kif9 is anchored to the nucleus and generates a pulling force that reels the centrosome up against the nucleus. This is a novel activity for a kinesin motor, one important for progression of cells into mitosis and to ensure centrosome-nuclear parity in a multinuclear environment.
doi:10.1007/s00018-012-1205-0
PMCID: PMC3594137  PMID: 23161062
Centrosome; Kinesin; Microtubule; Dictyostelium
4.  Kinetochore flexibility: creating a dynamic chromosome-spindle interface 
Kinetochores are complex macromolecular assemblies that link chromosomes to the mitotic spindle, mediate forces for chromosome motion, and generate the checkpoint signal delaying anaphase onset until all chromosomes are incorporated into the spindle. Proper execution of these functions depends on precise interactions between kinetochores and microtubules. While the molecular composition of the kinetochore is well described, structural organization of this organelle at the molecular and atomic levels is just beginning to emerge. Recent structural studies across scales suggest that kinetochores should not be viewed as rigid static scaffolds. Instead, these organelles exhibit a surprising degree of flexibility that enables rapid adaptations to various types of interactions with the mitotic spindle.
doi:10.1016/j.ceb.2011.12.008
PMCID: PMC3507511  PMID: 22221609
5.  Rules of engagement: centrosome–nuclear connections in a closed mitotic system 
Biology Open  2012;1(11):1111-1117.
Summary
The assembly of a functional mitotic spindle is essential for cell reproduction and requires a precise coordination between the nuclear cycle and the centrosome. This coordination is particularly prominent in organisms that undergo closed mitosis where centrosomes must not only respond to temporal signals, but also to spatial considerations, e.g. switching from the production of cytoplasmic microtubule arrays to the generation of dynamic intra-nuclear microtubules required for spindle assembly. We utilize a gene knockout of Kif9, a Dictyostelium discoideum Kin-I kinesin, to destabilize the physical association between centrosomes and the nuclear envelope. This approach presents a unique opportunity to reveal temporal and spatial components in the regulation of centrosomal activities in a closed-mitosis organism. Here we report that centrosome–nuclear engagement is not required for the entry into mitosis. Although detached centrosomes can duplicate in the cytoplasm, neither they nor nuclei alone can produce spindle-like microtubule arrays. However, the physical association of centrosomes and the nuclear envelope is required to progress through mitosis beyond prometaphase.
doi:10.1242/bio.20122188
PMCID: PMC3507195  PMID: 23213391
Centrosome; Nucleus; Closed mitosis; Dictyostelium
6.  The spatial arrangement of chromosomes during prometaphase facilitates spindle assembly 
Cell  2011;146(4):555-567.
Error-free chromosome segregation requires stable attachment of sister kinetochores to the opposite spindle poles (amphitelic attachment). Exactly how amphitelic attachments are achieved during spindle assembly remains elusive. We employed photoactivatable GFP and high-resolution live-cell confocal microscopy to visualize for the first time complete 3-D movements of individual kinetochores throughout mitosis in non-transformed human cells. Combined with electron microscopy, molecular perturbations, and immunofluorescence analyses, this approach reveals unexpected new details of chromosome behavior. Our data demonstrate that unstable lateral interactions between kinetochores and microtubules dominate during early prometaphase. These transient interactions lead to the reproducible arrangement of chromosomes in an equatorial ring on the surface of the nascent spindle. A computational model predicts that this toroidal distribution of chromosomes exposes kinetochores to a high-density of microtubules which facilitates subsequent formation of amphitelic attachments. Thus, spindle formation involves a previously overlooked stage of chromosome prepositioning which promotes formation of amphitelic attachments.
doi:10.1016/j.cell.2011.07.012
PMCID: PMC3291198  PMID: 21854981
mitosis; spindle assembly; chromosome congression; kinetochore
7.  Concerted effort of centrosomal and Golgi-derived microtubules is required for proper Golgi complex assembly but not for maintenance 
Molecular Biology of the Cell  2012;23(5):820-833.
Using computational modeling and laser microsurgery, we establish that neither the centrosomal microtubule array nor the Golgi-derived array is solely sufficient for correct Golgi assembly. Only the concerted effort of both MT arrays results in the integral, polarized Golgi complex necessary for polarized trafficking and cell motility.
Assembly of an integral Golgi complex is driven by microtubule (MT)-dependent transport. Conversely, the Golgi itself functions as an unconventional MT-organizing center (MTOC). This raises the question of whether Golgi assembly requires centrosomal MTs or can be self-organized, relying on its own MTOC activity. The computational model presented here predicts that each MT population is capable of gathering Golgi stacks but not of establishing Golgi complex integrity or polarity. In contrast, the concerted effort of two MT populations would assemble an integral, polarized Golgi complex. Indeed, while laser ablation of the centrosome did not alter already-formed Golgi complexes, acentrosomal cells fail to reassemble an integral complex upon nocodazole washout. Moreover, polarity of post-Golgi trafficking was compromised under these conditions, leading to strong deficiency in polarized cell migration. Our data indicate that centrosomal MTs complement Golgi self-organization for proper Golgi assembly and motile-cell polarization.
doi:10.1091/mbc.E11-06-0550
PMCID: PMC3290642  PMID: 22262454
8.  Centriole reduplication during prolonged interphase requires procentriole maturation governed by Plk1 
Current biology : CB  2010;20(14):1277-1282.
Summary
Supernumerary centrioles lead to abnormal mitosis [1,2] which in turn promotes tumorigenesis [3,4]. Thus, centriole duplication must be coordinated with the cell cycle to ensure that the number of centrioles in the cell doubles precisely during each cell cycle [5]. However, in some transformed cells centrioles undergo multiple rounds of duplication (reduplication) during prolonged interphase [6-8]. Mechanisms responsible for centriole reduplication are poorly understood. Here, we report that centrioles reduplicate consistently in cancerous and non-transformed human cells during G2 arrests and this reduplication requires the activity of Polo-like kinase 1 (Plk1). We also find that cell’s ability to reduplicate centrioles during S-arrests depends on the presence of activated (T210-phosphorylated) Plk1 at the centrosome. In the absence of activated Plk1, nascent procentrioles remain associated with mother centrioles, which prevent centriole reduplication. In contrast, if Plk1(pT210) appears at the centrosome, procentrioles mature, disengage from mother centrioles, and ultimately duplicate. Plk1 activity is not required for the assembly of procentrioles, however. Thus, the role of Plk1 is to coordinate centriole duplication cycle with the cell cycle. Activation of Plk1 during late-S-G2 induces procentriole maturation and after this point the centriole cycle can be completed autonomously, even in the absence of cell cycle progression.
doi:10.1016/j.cub.2010.05.050
PMCID: PMC2911792  PMID: 20656208
9.  Centriole duplication: analogue control in a digital age 
Cell biology international  2010;34(12):1239-1245.
In preparation for mitosis, the centrosome doubles once and only once to provide the two poles of the mitotic spindle. The presence of more than two centrosomes increases the chances that mitosis will be multipolar, and chromosomes will be distributed unequally. Since the number of mother–daughter centriole pairs determines the number of centrosomes, it is important that only one daughter centriole is assembled at, but slightly separated from, the proximal end of each mother centriole. This numerical and spatial specificity has led to the belief that a ‘template’ on the mother centriole provides a unique site for procentriole assembly. We review observations that are leading to the demise of this intuitively attractive idea. In its place, we are left with the notion that pericentriolar material at the wall of the mother centriole provides a local environment that promotes the assembly of a macromolecular complex that seeds the daughter centriole. Even though the system normally behaves in a digital fashion to go from zero to just one daughter centriole per mother, this behaviour appears to be based in the precise analogue control of multiple proteins, their activities, and the structure provided by the mother centriole.
doi:10.1042/CBI20100612
PMCID: PMC3051170  PMID: 21067522
cell cycle; centriole; centrosome; duplication; singularity; template
10.  Centromere tension: a divisive issue 
Nature cell biology  2010;12(10):919-923.
It has been proposed that the spindle assembly checkpoint detects both unattached kinetochores and lack of tension between sister kinetochores when sister chromatids are not attached to opposite spindle poles. However, here we argue that there is only one signal — whether kinetochores are attached to microtubules or not — and this has implications for our understanding of both chromosome segregation and the control of genomic stability.
doi:10.1038/ncb1010-919
PMCID: PMC3052926  PMID: 20885417
11.  The spindle position checkpoint requires positional feedback from cytoplasmic microtubules 
Current biology : CB  2009;19(23):2026-2030.
Summary
The objective of mitosis is to provide a copy of the genome to each progeny of a cell division. This requires the separation of duplicate chromatids by the spindle apparatus, and the delivery of one set of chromosomes to each of the daughter cells. In budding yeast, the fidelity of chromosome delivery depends on the spindle position checkpoint, which prolongs mitosis until one end of the anaphase spindle arrives in the bud. Here, we tested the hypothesis that the activity of the spindle position checkpoint depends on persistent interactions between cytoplasmic microtubules and the mother-bud neck, the future site of cytokinesis. We used laser ablation to disrupt microtubule interactions with the bud neck, and we found that loss of microtubules from the neck leads to mitotic exit in a majority of checkpoint-activated cells. Our findings suggest that cytoplasmic microtubules are used to monitor the location of the spindle in the dividing cell, and in the event of positioning errors, relay a signal to inhibit mitotic exit until the spindle is appropriately positioned.
doi:10.1016/j.cub.2009.10.020
PMCID: PMC2805762  PMID: 19913426
12.  Overly long centrioles and defective cell division upon excess of the SAS-4-related protein CPAP 
Current biology : CB  2009;19(12):1012-1018.
Summary
The centrosome is the principal microtubule organizing center (MTOC) of animal cells [1]. Accurate centrosome duplication is fundamental for genome integrity and entails the formation of one procentriole next to each existing centriole, once per cell cycle. The procentriole then elongates to eventually reach the same size as that of the centriole. The mechanisms that govern elongation of the centriolar cylinder and their potential relevance for cell division are not known. Here, we show that the SAS-4-related protein CPAP [2] is required for centrosome duplication in cycling human cells. Furthermore, we demonstrate that CPAP overexpression results in the formation of abnormal long centrioles. This also promotes formation of more than one procentriole in the vicinity of such overly long centrioles, eventually resulting in the presence of supernumerary MTOCs. This in turn leads to multipolar spindle assembly and cytokinesis defects. Overall, our findings suggest that centriole length must be carefully regulated to restrict procentriole number and thus ensure accurate cell division.
doi:10.1016/j.cub.2009.05.018
PMCID: PMC2993638  PMID: 19481460
13.  Mechanisms of chromosome behaviour during mitosis 
For over a century, scientists have strived to understand the mechanisms that govern the accurate segregation of chromosomes during mitosis. The most intriguing feature of this process, which is particularly prominent in higher eukaryotes, is the complex behaviour exhibited by the chromosomes. This behaviour is based on specific and highly regulated interactions between the chromosomes and spindle microtubules. Recent discoveries, enabled by high-resolution imaging combined with the various genetic, molecular, cell biological and chemical tools, support the idea that establishing and controlling the dynamic interaction between chromosomes and microtubules is a major factor in genomic fidelity.
doi:10.1038/nrm2832
PMCID: PMC2893392  PMID: 20068571
14.  The outer plate in vertebrate kinetochores is a flexible network with multiple microtubule interactions 
Nature cell biology  2007;9(5):516-522.
Intricate interactions between kinetochores and microtubules are essential for the proper distribution of chromosomes during mitosis. A crucial long-standing question is how vertebrate kinetochores generate chromosome motion while maintaining attachments to the dynamic plus ends of the multiple kinetochore MTs (kMTs) in a kinetochore fibre. Here, we demonstrate that individual kMTs in PtK1 cells are attached to the kinetochore outer plate by several fibres that either embed the microtubule plus-end tips in a radial mesh, or extend out from the outer plate to bind microtubule walls. The extended fibres also interact with the walls of nearby microtubules that are not part of the kinetochore fibre. These structural data, in combination with other recent reports, support a network model of kMT attachment wherein the fibrous network in the unbound outer plate, including the Hec1–Ndc80 complex, dissociates and rearranges to form kMT attachments.
doi:10.1038/ncb1576
PMCID: PMC2895818  PMID: 17435749
15.  Chromosome Congression in the Absence of K-Fibres 
Nature cell biology  2009;11(7):832-838.
Proper chromosome congression (the process of aligning chromosomes on the spindle) contributes to accurate and faithful chromosome segregation. It is widely accepted that congression requires ‘K-fibres’, microtubule bundles that extend from the kinetochores to spindle poles1, 2. Here we demonstrate that chromosomes in human cells co-depleted for HSET (kinesin-14)3, 4 and hNuf2 (a component of the Ndc80/Hec1 complex)5 can congress to the metaphase plate in the absence of K-fibres. However, the chromosomes were not stably maintained at the metaphase plate under these conditions. Chromosome congression in HSET+hNuf2 co-depleted cells required the plus-end directed motor CENP-E (kinesin-7)6, which has been implicated in the gliding of mono-oriented kinetochores alongside adjacent K-fibres7. Thus, proper end-on attachment of kinetochores to microtubules is not necessary for chromosome congression. Instead, our data support the idea that congression allows unattached chromosomes to move to the middle of the spindle where they have a higher probability of establishing connections with both spindle poles. These bi-oriented connections are also utilized to maintain stable chromosome alignment at the spindle equator.
doi:10.1038/ncb1890
PMCID: PMC2895821  PMID: 19525938
Chromosome congression; Kinetochore; K-fibre; Kinesin-14; hNuf2; HSET; CENP-E
16.  Mitosis: Too Much of a Good Thing (Can Be Bad) 
Current biology : CB  2009;19(22):R1032-R1034.
Recent studies reveal that the precise regulation of microtubule dynamics is essential for an error-free mitosis. Kinetochore microtubule attachments that are too stable increase the rate of chromosome mis-segregation, a leading cause of chromosomal instability in tumors.
doi:10.1016/j.cub.2009.10.013
PMCID: PMC2891095  PMID: 19948139
17.  The nature of cell-cycle checkpoints: facts and fallacies 
Journal of Biology  2009;8(10):88.
The concept of checkpoint controls revolutionized our understanding of the cell cycle. Here we revisit the defining features of checkpoints and argue that failure to properly appreciate the concept is leading to misinterpretation of experimental results. We illustrate, using the mitotic checkpoint, problems that can arise from a failure to respect strict definitions and precise terminology.
doi:10.1186/jbiol195
PMCID: PMC2790835  PMID: 19930621
18.  Relative contributions of chromatin and kinetochores to mitotic spindle assembly 
The Journal of Cell Biology  2009;187(1):43-51.
It's the kinetochores, not the DNA, that initiate spindle assembly.
During mitosis and meiosis in animal cells, chromosomes actively participate in spindle assembly by generating a gradient of Ran guanosine triphosphate (RanGTP). A high concentration of RanGTP promotes microtubule nucleation and stabilization in the vicinity of chromatin. However, the relative contributions of chromosome arms and centromeres/kinetochores in this process are not known. In this study, we address this issue using cells undergoing mitosis with unreplicated genomes (MUG). During MUG, chromatin is rapidly separated from the forming spindle, and both centrosomal and noncentrosomal spindle assembly pathways are active. MUG chromatin is coated with RCC1 and establishes a RanGTP gradient. However, a robust spindle forms around kinetochores/centromeres outside of the gradient peak. When stable kinetochore microtubule attachment is prevented by Nuf2 depletion in both MUG and normal mitosis, chromatin attracts astral microtubules but cannot induce spindle assembly. These results support a model in which kinetochores play the dominant role in the chromosome-mediated pathway of mitotic spindle assembly.
doi:10.1083/jcb.200903076
PMCID: PMC2762104  PMID: 19805628
19.  Ab ovo or de novo? Mechanisms of centriole duplication 
Molecules and cells  2009;27(2):135-142.
The centrosome, an organelle comprising centrioles and associated pericentriolar material, is the major microtubule organizing center in animal cells. For the cell to form a bipolar mitotic spindle and ensure proper chromosome segregation at the end of each cell cycle, it is paramount that the cell contains two and only two centrosomes. Because the number of centrosomes in the cell is determined by the number of centrioles, cells have evolved elaborate mechanisms to control centriole biogenesis and to tightly coordinate this process with DNA replication. Here we review key proteins involved in centriole assembly, compare two major modes of centriole biogenesis, and discuss the mechanisms that ensure stringency of centriole number.
doi:10.1007/s10059-009-0017-z
PMCID: PMC2691869  PMID: 19277494
centriole; centrosome; duplication; de novo
20.  hPOC5 is a centrin-binding protein required for assembly of full-length centrioles 
The Journal of Cell Biology  2009;185(1):101-114.
Centrin has been shown to be involved in centrosome biogenesis in a variety of eukaryotes. In this study, we characterize hPOC5, a conserved centrin-binding protein that contains Sfi1p-like repeats. hPOC5 is localized, like centrin, in the distal portion of human centrioles. hPOC5 recruitment to procentrioles occurs during G2/M, a process that continues up to the full maturation of the centriole during the next cell cycle and is correlated with hyperphosphorylation of the protein. In the absence of hPOC5, RPE1 cells arrest in G1 phase, whereas HeLa cells show an extended S phase followed by cell death. We show that hPOC5 is not required for the initiation of procentriole assembly but is essential for building the distal half of centrioles. Interestingly, the hPOC5 family reveals an evolutionary divergence between vertebrates and organisms like Drosophila melanogaster or Caenorhabditis elegans, in which the loss of hPOC5 may correlate with the conspicuous differences in centriolar structure.
doi:10.1083/jcb.200808082
PMCID: PMC2700515  PMID: 19349582
21.  Asymmetric CLASP-dependent nucleation of non-centrosomal microtubules at the trans-Golgi network 
Developmental cell  2007;12(6):917-930.
Proper organization of microtubule arrays is essential for intracellular trafficking and cell motility. It is generally assumed that most if not all microtubules in vertebrate somatic cells are formed by the centrosome. Here we demonstrate that a large number of microtubules in untreated human cells originate from the Golgi apparatus in a centrosome-independent manner. Both centrosomal and Golgi-emanating microtubules need γ-tubulin for nucleation. Additionally, formation of microtubules at the Golgi requires CLASPs, microtubule-binding proteins that selectively coat non-centrosomal microtubule seeds. We show that CLASPs are recruited to trans-Golgi network (TGN) at the Golgi periphery by the TGN protein GCC185. In sharp contrast to radial centrosomal arrays, microtubules nucleated at the peripheral Golgi compartment are preferentially oriented toward the leading edge in motile cells. We propose that Golgi–emanating microtubules contribute to the asymmetric microtubule networks in polarized cells and support diverse processes including post-Golgi transport to the cell front.
doi:10.1016/j.devcel.2007.04.002
PMCID: PMC2705290  PMID: 17543864
22.  The spindle assembly checkpoint is satisfied in the absence of interkinetochore tension during mitosis with unreplicated genomes 
The Journal of Cell Biology  2008;183(1):29-36.
The accuracy of chromosome segregation is enhanced by the spindle assembly checkpoint (SAC). The SAC is thought to monitor two distinct events: attachment of kinetochores to microtubules and the stretch of the centromere between the sister kinetochores that arises only when the chromosome becomes properly bioriented. We examined human cells undergoing mitosis with unreplicated genomes (MUG). Kinetochores in these cells are not paired, which implies that the centromere cannot be stretched; however, cells progress through mitosis. A SAC is present during MUG as cells arrest in response to nocodazole, taxol, or monastrol treatments. Mad2 is recruited to unattached MUG kinetochores and released upon their attachment. In contrast, BubR1 remains on attached kinetochores and exhibits a level of phosphorylation consistent with the inability of MUG spindles to establish normal levels of centromere tension. Thus, kinetochore attachment to microtubules is sufficient to satisfy the SAC even in the absence of interkinetochore tension.
doi:10.1083/jcb.200801038
PMCID: PMC2557037  PMID: 18824563
23.  Centrosome Amplification Can Initiate Tumorigenesis in Flies 
Cell  2008;133(6):1032-1042.
SUMMARY
Centrosome amplification is a common feature of many cancer cells, and it has been previously proposed that centrosome amplification can drive genetic instability and so tumorigenesis. To test this hypothesis, we generated Drosophila lines that have extra centrosomes in ~60% of their somatic cells. Many cells with extra centrosomes initially form multipolar spindles, but these spindles ultimately become bipolar. This requires a delay in mitosis that is mediated by the spindle assembly checkpoint (SAC). As a result of this delay, there is no dramatic increase in genetic instability in flies with extra centrosomes, and these flies maintain a stable diploid genome over many generations. The asymmetric division of the larval neural stem cells, however, is compromised in the presence of extra centrosomes, and larval brain cells with extra centrosomes can generate metastatic tumors when transplanted into the abdomens of wild-type hosts. Thus, centrosome amplification can initiate tumorigenesis in flies.
doi:10.1016/j.cell.2008.05.039
PMCID: PMC2653712  PMID: 18555779
24.  Peripheral mechanisms take central stage in microtubule dynamics: Commentary on "Signaling function of a-catenin in microtubule regulation" by Shtutman et al. 
Cell cycle (Georgetown, Tex.)  2008;7(15):2275.
Although the centrosome is traditionally viewed as cell’s principle microtubule organizing center (MTOC), regulation of microtubule dynamics at the cell cortex plays an equally important role in the formation of the steady-state microtubule network. Several recent studies, including one published in this issue, reveal that complex signaling mechanisms associated with adherence junctions influence both microtubule nucleation at the centrosome, and the stability of non-centrosomal microtubules.
PMCID: PMC2577214  PMID: 18682682
25.  Regulated HsSAS-6 Levels Ensure Formation of a Single Procentriole per Centriole during the Centrosome Duplication Cycle 
Developmental cell  2007;13(2):203-213.
SUMMARY
Centrosome duplication involves the formation of a single procentriole next to each centriole, once per cell cycle. The mechanisms governing procentriole formation and those restricting its occurrence to one event per centriole are poorly understood. Here, we show that HsSAS-6 is necessary for procentriole formation and that it localizes asymmetrically next to the centriole at the onset of procentriole formation. HsSAS-6 levels oscillate during the cell cycle, with the protein being degraded in mitosis and starting to accumulate again at the end of the following G1. Our findings indicate that APCCdh1 targets HsSAS-6 for degradation by the 26S proteasome. Importantly, we demonstrate that increased HsSAS-6 levels promote formation of more than one procentriole per centriole. Therefore, regulated HsSAS-6 levels normally ensure that each centriole seeds the formation of a single procentriole per cell cycle, thus playing a fundamental role in driving the centrosome duplication cycle and ensuring genome integrity.
doi:10.1016/j.devcel.2007.07.004
PMCID: PMC2628752  PMID: 17681132

Results 1-25 (50)