Site Of Cytokinesis Research Articles

Microtubule reorganization during mitotic cell division in the dinoflagellate Ostreospis cf. ovata.

Dinoflagellates are marine organisms that undergo seasonal proliferation events known as algal blooms. Vegetative cell proliferation is a main contributing factor in these events. However, mechanistical understanding of mitosis and cytokinesis in dinoflagellates remains rudimentary. Using an optimized immunofluorescence protocol, we analysed changes in microtubule organization occurring during the mitotic cycle of the toxic dinoflagellate Ostreopsis cf. ovata. We find that the flagella and the cortical microtubule array persist throughout the mitotic cycle. Two cytoplasmic microtubule bundles originate from the ventral area, where the basal bodies are located - a cortical bundle and a cytoplasmic bundle. The latter associates with the nucleus in the cell centre before mitosis and with the acentrosomal extranuclear spindle during mitosis. Analysis of tubulin post-translational modifications identifies two populations of spindle microtubules - polar acetylated microtubules, whose length is constant, and central tyrosinated microtubules, which elongate during chromosome segregation. During cell division a microtubule-rich structure forms along the dorsal-ventral axis, associated with the site of cytokinesis, consistent with a cytokinetic mechanism that is independent of the actomyosin ring typical of animal and yeast cells.

Simultaneous co-overexpression of Saccharomyces cerevisiae septins Cdc3 and Cdc10 drives pervasive, phospholipid-, and tag-dependent plasma membrane localization.

Septin proteins contribute to many eukaryotic processes involving cellular membranes. In the budding yeast Saccharomyces cerevisiae, septin hetero-oligomers interact with the plasma membrane (PM) almost exclusively at the future site of cytokinesis. While multiple mechanisms of membrane recruitment have been identified, including direct interactions with specific phospholipids and curvature-sensitive interactions via amphipathic helices, these do not fully explain why yeast septins do not localize all over the inner leaflet of the PM. While engineering an inducible split-yellow fluorescent protein (YFP) system to measure the kinetics of yeast septin complex assembly, we found that ectopic co-overexpression of two tagged septins, Cdc3 and Cdc10, resulted in nearly uniform PM localization, as well as perturbation of endogenous septin function. Septin localization and function in gametogenesis were also perturbed. PM localization required the C-terminal YFP fragment fused to the C terminus of Cdc3, the septin-associated kinases Cla4 and Gin4, and phosphotidylinositol-4,5-bis-phosphate (PI[4,5]P2 ), but not the putative PI(4,5)P2 -binding residues in Cdc3. Endogenous Cdc10 was recruited to the PM, likely contributing to the functional interference. PM-localized septins did not exchange with the cytosolic pool, indicative of stable polymers. These findings provide new clues as to what normally restricts septin localization to specific membranes.

Microtubule organization and plastid distribution during meiosis of Haplomitrium mnioides (Haplomitriopsida)

The organization of microtubules and plastid distribution of the liverwort, Haplomitrium mnioides (Haplomitriopsida), was studied during the meiotic phase lasting for six months. In the late fall, the cytoplasm of early sporocytes forms four lobes of future spore domains before meiotic prophase. Microtubules align at the cytoplasmic cleavage furrow regions as girdling bands in the four-lobed sporocytes. Finally, the cleavage furrows are proximal to the nucleus positioned in the center of the sporocyte, and the girdling bands of the microtubule (GBM) disappear. Subsequently, the nucleus moves into one of the cytoplasmic lobes, and sporocytes pass the winter season at this stage. In early spring, the nucleus returns to the central position of the lobed cytoplasm, concurrent with plastid repositioning around the nucleus. Plastids are then distributed equally to each of the four lobes as a plastid cluster. Astral microtubules emanate from the plastid cluster in each spore domain and encage prophase nuclei as a quadripolar microtubule system (QMS). The QMS changes into a twisted spindle of metaphase I with broad poles, while spindles of metaphase II also emanate from the four plastid clusters. Cytokinesis is completed through the centrifugal cell plate formation in telophase II. The division axes of two successive nuclear divisions appear to be determined by plastid-based QMS, and the future site of cytokinesis is marked by cytoplasmic furrows associated with GBM. The phylogenetic distribution of GBM and QMS suggests that the meiotic system involving these structures is an ancestral trait of liverworts. Long-term dormancy in diploid sporocytes rather than haploid spores may represent transitional traits from charophycean green algae to land plants.

A cell separation checkpoint that enforces the proper order of late cytokinetic events.

Eukaryotic cell division requires dependency relationships in which late processes commence only after early ones are appropriately completed. We have discovered a system that blocks late events of cytokinesis until early ones are successfully accomplished. In budding yeast, cytokinetic actomyosin ring contraction and membrane ingression are coupled with deposition of an extracellular septum that is selectively degraded in its primary septum immediately after its completion by secreted enzymes. We find this secretion event is linked to septum completion and forestalled when the process is slowed. Delay of septum degradation requires Fir1, an intrinsically disordered protein localized to the cytokinesis site that is degraded upon septum completion but stabilized when septation is aberrant. Fir1 protects cytokinesis in part by inhibiting a separation-specific exocytosis function of the NDR/LATS kinase Cbk1, a key component of "hippo" signaling that induces mother-daughter separation. We term this system enforcement of cytokinesis order, a checkpoint ensuring proper temporal sequence of mechanistically incompatible processes of cytokinesis.

Cytokinesis requires localized \u03b2-actin filament production by an actin isoform specific nucleator

Cytokinesis is initiated by the localized assembly of the contractile ring, a dynamic actomyosin structure that generates a membrane furrow between the segregating chromosomal masses to divide a cell into two. Here we show that the stabilization and organization of the cytokinetic furrow is specifically dependent on localized β-actin filament assembly at the site of cytokinesis. β-actin filaments are assembled directly at the furrow by an anillin-dependent pathway that enhances RhoA-dependent activation of the formin DIAPH3, an actin nucleator. DIAPH3 specifically generates homopolymeric filaments of β-actin in vitro. By employing enhancers and activators, cells can achieve acute spatio-temporal control over isoform-specific actin arrays that are required for distinct cellular functions.

Dynein is regulated by the stability of its microtubule track.

How dynein motors accurately move cargoes is an important question. In budding yeast, dynein moves the mitotic spindle to the predetermined site of cytokinesis by pulling on astral microtubules. In this study, using high-resolution imaging in living cells, we discover that spindle movement is regulated by changes in microtubule plus-end dynamics that occur when dynein generates force. Mutants that increase plus-end stability increase the frequency and duration of spindle movements, causing positioning errors. We find that dynein plays a primary role in regulating microtubule dynamics by destabilizing microtubules. In contrast, the dynactin complex counteracts dynein and stabilizes microtubules through a mechanism involving the shoulder subcomplex and the cytoskeletal-associated protein glycine-rich domain of Nip100/p150glued Our results support a model in which dynein destabilizes its microtubule substrate by using its motility to deplete dynactin from the plus end. We propose that interplay among dynein, dynactin, and the stability of the microtubule substrate creates a mechanism that regulates accurate spindle positioning.

Chromatin and Cell Wall Staining of Schizosaccharomyces pombe.

Fission yeasts grow by tip extension, maintaining a constant width until they reach a critical size threshold and divide. Division by medial fission-which gives these yeast their name-generates a new end that arises from the site of cytokinesis. The old end, which was produced during the previous cell cycle, initiates progression of the new cell cycle, and in G2, the new end is activated in a process termed new-end takeoff (NETO). In this protocol, the fluorescent stains calcofluor and 4',6-diamidino-2-phenylindole (DAPI) are used to give a rapid and informative assessment of morphogenesis and cell-cycle progression in the fission yeast Schizosaccharomyces pombe Calcofluor reveals the timing of NETO because it stains the birth scars that are generated at new ends by cytokinesis less efficiently than the rest of the cell wall. Intense calcofluor staining of the septum and measurement of cell length are also widely used to identify dividing cells and to gauge the timing of mitotic commitment. Staining nuclei with DAPI identifies mono- and binucleated cells and complements the calcofluor staining procedure to evaluate the stages of the cell cycle and identify mitotic errors. Equally simple DAPI staining procedures reveal chromatin structure in higher resolution, facilitating more accurate staging of mitotic progression and characterization of mitotic errors.

Splitsville: structural and functional insights into the dynamic bacterial Z ring.

Bacteria must divide to increase in number and colonize their niche. Binary fission is the most widespread means of bacterial cell division, but even this relatively simple mechanism has many variations on a theme. In most bacteria, the tubulin homologue FtsZ assembles into a ring structure, termed the Z ring, at the site of cytokinesis and recruits additional proteins to form a large protein machine - the divisome - that spans the membrane. In this Review, we discuss current insights into the regulation of the assembly of the Z ring and how the divisome drives membrane invagination and septal cell wall growth while flexibly responding to various cellular inputs.

Comprehensive Genetic Analysis of Paralogous Terminal Septin Subunits Shs1 and Cdc11 in Saccharomyces cerevisiae.

Septins are a family of GTP-binding proteins considered to be cytoskeletal elements because they self-assemble into filaments and other higher-order structures in vivo. In budding yeast, septins establish a diffusion barrier at the bud neck between a mother and daughter cell, promote membrane curvature there, and serve as a scaffold to recruit other proteins to the site of cytokinesis. However, the mechanism by which any septin engages a partner protein has been unclear. The two most related and recently evolved subunits appear to be Cdc11 and Shs1, and the basic building blocks for assembling septin structures are hetero-octameric rods (Cdc11-Cdc12-Cdc3-Cdc10-Cdc10-Cdc3-Cdc12-Cdc11 and Shs1-Cdc12-Cdc3-Cdc10-Cdc10-Cdc3-Cdc12-Shs1). Loss of Cdc11 is not normally tolerated, whereas cells lacking Shs1 do not appear grossly abnormal. We established several different sensitized genetic backgrounds wherein Shs1 is indispensable, which allowed us to carry out the first comprehensive and detailed genetic analysis of Shs1 in vivo. Our analysis revealed several novel insights, including: (i) the sole portion of Shs1 essential for its function is a predicted coiled-coil-forming segment in its C-terminal extension (CTE); (ii) the CTE of Cdc11 shares this function; (iii) this role for the CTEs of Cdc11 and Shs1 is quite distinct from that of the CTEs of Cdc3 and Cdc12; and (iv) heterotypic Cdc11 and Shs1 junctions likely occur in vivo.

Interdomain allosteric regulation of Polo kinase by Aurora B and Map205 is required for cytokinesis.

Drosophila melanogaster Polo and its human orthologue Polo-like kinase 1 fulfill essential roles during cell division. Members of the Polo-like kinase (Plk) family contain an N-terminal kinase domain (KD) and a C-terminal Polo-Box domain (PBD), which mediates protein interactions. How Plks are regulated in cytokinesis is poorly understood. Here we show that phosphorylation of Polo by Aurora B is required for cytokinesis. This phosphorylation in the activation loop of the KD promotes the dissociation of Polo from the PBD-bound microtubule-associated protein Map205, which acts as an allosteric inhibitor of Polo kinase activity. This mechanism allows the release of active Polo from microtubules of the central spindle and its recruitment to the site of cytokinesis. Failure in Polo phosphorylation results in both early and late cytokinesis defects. Importantly, the antagonistic regulation of Polo by Aurora B and Map205 in cytokinesis reveals that interdomain allosteric mechanisms can play important roles in controlling the cellular functions of Plks.

A Safeguard Mechanism Regulates Rho GTPases to Coordinate Cytokinesis with the Establishment of Cell Polarity

Author SummaryIn budding yeast, cell polarization (or the asymmetric distribution of subcellular components) ensures the targeted transport of proteins and membrane material to the sites of cell growth or cell division in late mitosis. Two conserved members of the Rho-GTPase family, Rho1 and Cdc42, are master regulators of cell polarity. While Rho1 has a well-established role in cytokinesis and cell separation, Cdc42 helps to establish the new polarity site from which the future daughter cell will grow after cytokinesis. Interestingly, despite the fact that Cdc42 is recruited to the site of cell division at the same time as Rho1, the new daughter cell never emerges from the site previously used for cytokinesis during the preceding cell cycle, and it remains elusive how cells coordinate the distinct functions of Rho1 and Cdc42 during cytokinesis. Here, we show that the novel protein Gps1 marks the cell division site, where it maintains Rho1-dependent polarity until cell separation is completed. We also demonstrate that Gps1 prevents activation of Cdc42 at the site of cell division during cytokinesis. We propose that Gps1 provides a novel polarity cue that guides the establishment of a new polarity site, away from the old site of cell division, where the new daughter cell then emerges.

Polarization of Diploid Daughter Cells Directed by Spatial Cues and GTP Hydrolysis of Cdc42 in Budding Yeast

Cell polarization occurs along a single axis that is generally determined by a spatial cue. Cells of the budding yeast exhibit a characteristic pattern of budding, which depends on cell-type-specific cortical markers, reflecting a genetic programming for the site of cell polarization. The Cdc42 GTPase plays a key role in cell polarization in various cell types. Although previous studies in budding yeast suggested positive feedback loops whereby Cdc42 becomes polarized, these mechanisms do not include spatial cues, neglecting the normal patterns of budding. Here we combine live-cell imaging and mathematical modeling to understand how diploid daughter cells establish polarity preferentially at the pole distal to the previous division site. Live-cell imaging shows that daughter cells of diploids exhibit dynamic polarization of Cdc42-GTP, which localizes to the bud tip until the M phase, to the division site at cytokinesis, and then to the distal pole in the next G1 phase. The strong bias toward distal budding of daughter cells requires the distal-pole tag Bud8 and Rga1, a GTPase activating protein for Cdc42, which inhibits budding at the cytokinesis site. Unexpectedly, we also find that over 50% of daughter cells lacking Rga1 exhibit persistent Cdc42-GTP polarization at the bud tip and the distal pole, revealing an additional role of Rga1 in spatiotemporal regulation of Cdc42 and thus in the pattern of polarized growth. Mathematical modeling indeed reveals robust Cdc42-GTP clustering at the distal pole in diploid daughter cells despite random perturbation of the landmark cues. Moreover, modeling predicts different dynamics of Cdc42-GTP polarization when the landmark level and the initial level of Cdc42-GTP at the division site are perturbed by noise added in the model.

A kinetoplastid‐specific kinesin is required for cytokinesis and for maintenance of cell morphology in Trypanosoma brucei

Kinesins are motor-based transport proteins that play diverse roles in various cellular processes. The trypanosome genome lacks the homologues of many conserved mitotic kinesins, but encodes a number of trypanosome-specific kinesins with unknown function. Here, we report the biochemical and functional characterization of TbKIN-C, a trypanosome-specific kinesin, which was initially identified through an RNAi screen for cytokinesis genes in T. brucei. TbKIN-C possesses in vitro ATPase activity and associates with cytoskeletal tubulin microtubules in vivo. It is distributed throughout the cytoskeleton with a focal enrichment at the posterior end of the cell during early cell cycle stages. RNAi of TbKIN-C resulted in distorted cell shape with an elongated posterior filled with tyrosinated tubulin microtubules. Silencing of TbKIN-C impaired the segregation of organelles and cytoskeletal structures and led to detachment of the new flagellum and a small portion of the cytoplasm. We also show that RNAi of TbKIN-C compromised cytokinesis and abolished the trans-localization of TbCPC1, a subunit of the chromosomal passenger complex, from the central spindle to the initiation site of cytokinesis. Our results suggest an essential role of TbKIN-C in maintaining cell morphology, likely through regulating microtubule dynamics at the posterior end of the cell.

At the Right Time and at the Right Place: Control of Cytokinesis in Mammals

Ensuring that cytokinesis occurs at the right time and in the right place is essential to maintain euploidy and cell-cycle progression. Cytokinesis is triggered by the RhoA GEF Ect2, which localizes to the spindle midzone during anaphase and activates RhoA at the overlying cell cortex to stimulate contractile ring formation. Using a chemical inhibitor of the Polo kinase Plk1, Petronczki et al. (2007) showed that Plk1 is essential for cytokinesis, because it specifically promotes Ect2 localization and interaction with its midzone receptor HsCyk-4. I liked this paper for three reasons. As a biologist, I was impressed because the paper explains an important part of biology: how a cell-cycle kinase determines when and where cytokinesis occurs. As a geneticist, I appreciate a tight allele. The paper is a beautiful example of the power of chemical genetics to specifically and rapidly inhibit Polo at a specific cell-cycle stage. Finally, as a yeast cell-cycle researcher, it is always nice to see the dramatic conservation between yeast and mammals. In budding yeast, even though the bud neck and not the spindle midzone determines the site of cytokinesis, Polo kinase phosphorylates Rho GEFs to initiate contractile ring formation and cytokinesis. This PaperPick refers to Polo-like Kinase 1 Triggers the Initiation of Cytokinesis in Human Cells by Promoting Recruitment of the RhoGEF Ect2 to the Central Spindle, by M. Petronczki, M. Glotzer, N. Kraut, and J.-M. Peters, published in May 2007. Video Abstract Here, Jan-Michael Peters discusses the work as a collaboration between Mark Petronczki (in his group), Michael Glotzer of the University of Chicago, and Norbert Kraut of Boehringer Ingelheim, who provided the key reagent: BI 2536, a chemical inhibitor of Plk1. The authors would like to note that their work on the role of Plk1 in cytokinesis was inspired by early observations made in the laboratory of David Glover and that the research groups of Prasad Jallepalli (Sloan Kettering) and Aaron Straight (Stanford) have also identified Plk1 as a key regulator of cleavage furrow formation in mammalian cells.

Spatial uncoupling of mitosis and cytokinesis during appressorium-mediated plant infection by the rice blast fungus Magnaporthe oryzae.

To infect plants, many pathogenic fungi develop specialized infection structures called appressoria. Here, we report that appressorium development in the rice blast fungus Magnaporthe oryzae involves an unusual cell division, in which nuclear division is spatially uncoupled from the site of cytokinesis and septum formation. The position of the appressorium septum is defined prior to mitosis by formation of a heteromeric septin ring complex, which was visualized by spatial localization of Septin4:green fluorescent protein (GFP) and Septin5:GFP fusion proteins. Mitosis in the fungal germ tube is followed by long-distance nuclear migration and rapid formation of an actomyosin contractile ring in the neck of the developing appressorium, at a position previously marked by the septin complex. By contrast, mutants impaired in appressorium development, such as Deltapmk1 and DeltacpkA regulatory mutants, undergo coupled mitosis and cytokinesis within the germ tube. Perturbation of the spatial control of septation, by conditional mutation of the SEPTATION-ASSOCIATED1 gene of M. oryzae, prevented the fungus from causing rice blast disease. Overexpression of SEP1 did not affect septation during appressorium formation, but instead led to decoupling of nuclear division and cytokinesis in nongerminated conidial cells. When considered together, these results indicate that SEP1 is essential for determining the position and frequency of cell division sites in M. oryzae and demonstrate that differentiation of appressoria requires a cytokinetic event that is distinct from cell divisions within hyphae.

The Spindle Position Checkpoint Requires Positional Feedback from Cytoplasmic Microtubules

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.

Trafficking of Crumbs3 during Cytokinesis Is Crucial for Lumen Formation

Although lumen generation has been extensively studied through so-called cyst-formation assays in Madin-Darby canine kidney (MDCK) cells, an underlying mechanism that leads to the initial appearance of a solitary lumen remains elusive. Lumen formation is thought to take place at early stages in aggregates containing only a few cells. Evolutionarily conserved polarity protein complexes, namely the Crumbs, Par, and Scribble complexes, establish apicobasal polarity in epithelial cells, and interference with their function impairs the regulated formation of solitary epithelial lumina. Here, we demonstrate that MDCK cells form solitary lumina during their first cell division. Before mitosis, Crumbs3a becomes internalized and concentrated in Rab11-positive recycling endosomes. These compartments become partitioned in both daughter cells and are delivered to the site of cytokinesis, thus forming the first apical membrane, which will eventually form a lumen. Endosome trafficking in this context appears to depend on the mitotic spindle apparatus and midzone microtubules. Furthermore, we show that this early lumen formation is regulated by the apical polarity complexes because Crumbs3 assists in the recruitment of aPKC to the forming apical membrane and interference with their function can lead to the formation of a no-lumen or multiple-lumen phenotype at the two-cell stage.

Binding of PI4,5P2 by septin complexes is required for their essential function in cytokinesis in budding yeast

Septins comprise a conserved family of proteins that are involved in processes of membrane remodeling, such as cytokinesis. Cytokinesis in budding yeast (Saccharomyces cerevisiae) requires assembly of septins adjacent to the plasma membrane of the mother‐bud neck, where they serve as a protein scaffold and diffusion barrier. The aim of our studies is to determine how septins are recruited to and anchored at the plasma membrane. Among plasma membrane constituents, phosphatidylinositol‐4,5‐bisphosphate (PI4,5P2) is a potential candidate for recruiting septins, because at the site of cytokinesis, the plasma membrane in mammalian cells is enriched in PI4,5P2, which is required for cytokinesis [Emoto et al. (2005) J. Biol. Chem. 280: 37901]. PI4,5P2 is also required for proper septin localization in yeast [Rodríguez‐Escudero et al. (2005) Biochem. J. 390: 613]. Our work indicates that a PI4,5P2‐binding reporter localizes preferentially to the yeast bud neck. In addition, liposome flotation assays show that the yeast septins Cdc3, Cdc11 and Cdc12 bind to PI4,5P2‐containing vesicles in vitro. Furthermore, complexes of Cdc3, Cdc10, Cdc11 and Cdc12 also bind PI4,5P2. Mutation of basic residues in Cdc3 decreases PI4,5P2 binding in vitro and causes defects in cytokinesis in vivo, supporting the view that septin‐PI4,5P2 interaction is required for the septins to execute their function in cytokinesis. NIH R01 GM21841

Polo-Like Kinase Is Expressed in S/G 2 /M Phase and Associated with the Flagellum Attachment Zone in both Procyclic and Bloodstream Forms of Trypanosoma brucei

Trypanosoma brucei, the etiologic agent of African sleeping sickness, divides into insect (procyclic) and bloodstream forms. These two forms are subject to distinct cell cycle regulations, with cytokinesis controlled primarily by basal body/kinetoplast segregation in the procyclic form but by mitosis in the bloodstream form. Polo-like kinases (PLKs), known to play essential roles in regulating both mitosis and cytokinesis among eukaryotes, have a homologue in T. brucei, TbPLK, which regulates only cytokinesis. In our previous study, overexpressed triply hemagglutinin-tagged TbPLK (TbPLK-3HA) in the procyclic form localized to a mid-dorsal point and the anterior tip of the cell along the flagellum attachment zone (FAZ). In our current study, TbPLK-3HA expressed at the endogenous level was identified at the same dorsal location of both procyclic and bloodstream forms, albeit it was no longer detectable at the anterior tip of the cell. Endogenously expressed TbPLK fused with an enhanced yellow fluorescent protein (EYFP) localized to the same dorsal location along the FAZs in living procyclic and bloodstream cells. Fluorescence-activated cell sorter analysis of hydroxyurea-synchronized procyclic cells revealed that TbPLK-EYFP emerges during S phase, persists through G(2)/M phase, and vanishes in G(1) phase. An indicated TbPLK-EYFP association with the FAZs of G(2)/M cells may thus represent a timely localization to a potential initiation site of cytokinesis, which agrees with the recognized role of TbPLK in cytokinetic initiation.

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Ensuring that cytokinesis occurs at the right time and in the right place is essential to maintain euploidy and cell-cycle progression. Cytokinesis is triggered by the RhoA GEF Ect2, which localizes to the spindle midzone during anaphase and activates RhoA at the overlying cell cortex to stimulate contractile ring formation. Using a chemical inhibitor of the Polo kinase Plk1, Petronczki et al. (2007) showed that Plk1 is essential for cytokinesis, because it specifically promotes Ect2 localization and interaction with its midzone receptor HsCyk-4.I liked this paper for three reasons. As a biologist, I was impressed because the paper explains an important part of biology: how a cell-cycle kinase determines when and where cytokinesis occurs. As a geneticist, I appreciate a “tight allele.” The paper is a beautiful example of the power of chemical genetics to specifically and rapidly inhibit Polo at a specific cell-cycle stage. Finally, as a yeast cell-cycle researcher, it is always nice to see the dramatic conservation between yeast and mammals. In budding yeast, even though the bud neck and not the spindle midzone determines the site of cytokinesis, Polo kinase phosphorylates Rho GEFs to initiate contractile ring formation and cytokinesis.This PaperPick refers to “Polo-like Kinase 1 Triggers the Initiation of Cytokinesis in Human Cells by Promoting Recruitment of the RhoGEF Ect2 to the Central Spindle,” by M. Petronczki, M. Glotzer, N. Kraut, and J.-M. Peters, published in May 2007.