Localization of Pavarotti-KLP in Living<i>Drosophila</i>Embryos Suggests Roles in Reorganizing the Cortical Cytoskeleton during the Mitotic Cycle

Minestrini, Gianluca; Harley, Alyssa Skye; Glover, David M.

doi:10.1091/mbc.e03-04-0214

Cited by 77 publications

(93 citation statements)

References 29 publications

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“…Because the actin cytoskeleton is in the cortex and Arfs are involved in both membrane trafficking and actin dynamics (Norman et al, 1998;Donaldson, 2003), association between the midbody and the cell cortex could be mediated through the actin/Arfbinding domain of CHO1. In support of this hypothesis, PAV-KLP has recently been demonstrated to locate at the cortical equatorial ring during cytokinesis in Drosophila cells (Somers and Saint, 2003;Minestrini et al, 2003). We also detected the interaction of CHO1 with the cortex in cytochalasin D-treated mitotic CHO cells (unpublished results).…”

Section: Role Of T1 and T2 Subregions Of The Tailsupporting

confidence: 78%

“…Although PAV-KLP in Drosophila has a functional NLS, corresponding to NLS1 in vertebrates, nuclear localization of the motor protein appears to be essential only for the female germ line development (Minestrini et al, 2002), but not for embryonic cell division (Minestrini et al, 2003). In C. elegans, ZEN-4 lacks not only NLS but also the entire C-terminus corresponding to the NLS-containing region.…”

Section: Role Of the Nls-containing Regionmentioning

confidence: 99%

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Role of the Midbody Matrix in Cytokinesis: RNAi and Genetic Rescue Analysis of the Mammalian Motor Protein CHO1

Matulienė

Kuriyama

2004

MBoC

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CHO1 is a kinesin-like motor protein essential for cytokinesis in mammalian cells. To analyze how CHO1 functions, we established RNAi and genetic rescue assays. CHO1-depleted cells reached a late stage of cytokinesis but fused back to form binucleate cells because of the absence of the midbody matrix in the middle of the intercellular bridge. Expression of exogenous CHO1 restored the formation of the midbody matrix and rescued cytokinesis in siRNA-treated cells. By analyzing phenotypes rescued with different constructs, it was shown that both motor and stalk domains function in midbody formation, whereas the tail is essential for completion of cytokinesis after the midbody matrix has formed. During the terminal stage of cytokinesis, different subregions of the tail play distinctive roles in stabilizing the midbody matrix and maintaining an association between the midbody and cell cortex. These results demonstrate that CHO1 consists of functionally differentiated subregions that act in concert to ensure complete cell separation.

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Section: Role Of T1 and T2 Subregions Of The Tailsupporting

confidence: 78%

Section: Role Of the Nls-containing Regionmentioning

confidence: 99%

Role of the Midbody Matrix in Cytokinesis: RNAi and Genetic Rescue Analysis of the Mammalian Motor Protein CHO1

Matulienė

Kuriyama

2004

MBoC

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“…This similar phenotype of the centralspindlin complex and the F-actin affecting proteins suggests that centralspindlin may be an upstream regulator of F-actin filament formation. Indeed kinase-dead mutants of pavarotti have been shown to accumulate at the spindle poles and are associated with an abnormal accumulation of F-actin near the centrosomes (48). Centralspindlin may be acting indirectly by helping to localize an important actinaffecting protein at the central spindle, or it may act more directly on the cortex.…”

Section: Contraction Of Actomyosin Structures During Cell Division Anmentioning

confidence: 99%

Distinct pathways control recruitment and maintenance of myosin II at the cleavage furrow during cytokinesis

Dean

Rogers

Stuurman

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

106

101

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The correct localization of myosin II to the equatorial cortex is crucial for proper cell division. Here, we examine a collection of genes that cause defects in cytokinesis and reveal with live cell imaging two distinct phases of myosin II localization. Three genes in the rho1 signaling pathway, pebble (a Rho guanidine nucleotide exchange factor), rho1, and rho kinase, are required for the initial recruitment of myosin II to the equatorial cortex. This initial localization mechanism does not require F-actin or the two components of the centralspindlin complex, the mitotic kinesin pavarotti͞MKLP1 and racGAP50c͞CYK-4. However, F-actin, the centralspindlin complex, formin (diaphanous), and profilin (chickadee) are required to stably maintain myosin II at the furrow. In the absence of these latter genes, myosin II delocalizes from the equatorial cortex and undergoes highly dynamic appearances and disappearances around the entire cell cortex, sometimes associated with abnormal contractions or blebbing. Our findings support a model in which a rho kinase-dependent event, possibly myosin II regulatory light chain phosphorylation, is required for the initial recruitment to the furrow, whereas the assembly of parallel, unbranched actin filaments, generated by formin-mediated actin nucleation, is required for maintaining myosin II exclusively at the equatorial cortex.contractile ring ͉ rho1

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“…In mammalian cells, MKLP1 is required for the formation of the midbody matrix and the completion of cytokinesis (7,8). Similar gene disruption experiments of MKLP1 orthologs in zebrafish (9), Drosophila (10), and Caenorhabditis elegans (11) also result in a cytokinesis failure. In most cases, furrow ingression is initiated, but cytokinesis fails to complete.…”

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confidence: 92%

A mitotic kinesin-like protein required for normal karyokinesis, myosin localization to the furrow, and cytokinesis in Dictyostelium

Lakshmikanth

Warrick²,

Spudich³

2004

Proc. Natl. Acad. Sci. U.S.A.

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Dictyostelium mitotic kinesin Kif12 is required for cytokinesis. Myosin II localization to the cleavage furrow is severely depressed in Kif12-null (⌬kif12) cells, which accounts in part for the cytokinesis failure. Myosin II-null cells, however, undergo mitosis-coupled cytokinesis when adhering to a surface, whereas the ⌬kif12 cells cannot. During mitosis, the rate of change of internuclear separation in ⌬kif12 cells is reduced compared with wild-type cells, indicating multiple roles of this molecular motor during mitosis and cytokinesis. GFP-Kif12, which rescues wild-type behavior when expressed in the ⌬kif12 strain, is concentrated in the nucleus in interphase cells, translocates to the cytoplasm at the onset of mitosis, appears in the centrosomes and spindle, and later is concentrated in the spindle midbody. Given these results, we hypothesize a mechanism for myosin II translocation to the furrow to set up the contractile ring.Kif12 ͉ cell-cycle regulation ͉ mitosis C ytokinesis and karyokinesis are fundamental to cell division and propagation. Karyokinesis involves nuclear segregation and requires the formation of a microtubule network on which the chromosomes are segregated with the help of kinesin-based motors. Cytokinesis involves division of cellular components of a parental cell into two daughter cells, and it is precisely timed along with nuclear division in almost all cells. Cytokinesis is generally achieved by an actin-myosin-based contractile ring that is assembled along the plane perpendicular to the axis of the spindle poles. Karyokinesis and cytokinesis are tightly synchronized to ensure high fidelity of genomic information transfer.Classic experiments carried out on sand dollar eggs established the importance of the mitotic spindles and astral microtubules in setting up the contractile ring (1, 2). The central spindle has been shown to be important for the completion of cytokinesis in several organisms, including Dictyostelium (3). Although a number of proteins have been shown to be associated with the mitotic spindle and implicated in cytokinesis (4-6), the molecular clues for the crosstalk between the spindle and cortical midzone during mitosis remain elusive.Members of the mitotic kinesin family have been shown to play key roles in the assembly and function of the mitotic spindle. In mammalian cells, MKLP1 is required for the formation of the midbody matrix and the completion of cytokinesis (7,8). Similar gene disruption experiments of MKLP1 orthologs in zebrafish (9), Drosophila (10), and Caenorhabditis elegans (11) also result in a cytokinesis failure. In most cases, furrow ingression is initiated, but cytokinesis fails to complete. Thus, MKLP1 is a key player in passing information from the mitotic spindle to the cleavage furrow.Myosin II is a major component of the cellular machinery responsible for cytokinesis (12, 13). However, little is known about how myosin is translocated to the cleavage furrow during mitosis. Using Dictyostelium as a model system, our laboratory has been studying m...

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Localization of Pavarotti-KLP in LivingDrosophilaEmbryos Suggests Roles in Reorganizing the Cortical Cytoskeleton during the Mitotic Cycle

Cited by 77 publications

References 29 publications

Role of the Midbody Matrix in Cytokinesis: RNAi and Genetic Rescue Analysis of the Mammalian Motor Protein CHO1

Role of the Midbody Matrix in Cytokinesis: RNAi and Genetic Rescue Analysis of the Mammalian Motor Protein CHO1

Distinct pathways control recruitment and maintenance of myosin II at the cleavage furrow during cytokinesis

A mitotic kinesin-like protein required for normal karyokinesis, myosin localization to the furrow, and cytokinesis in Dictyostelium

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