The Schizosaccharomyces pombe actin-related protein, Arp3, is a component of the cortical actin cytoskeleton and interacts with profilin.

McCollum, Dannel; Feoktistova, Anna; Morphew, Mary K.; Balasubramanian, Mohan K.; Gould, Kathleen L.

doi:10.1002/j.1460-2075.1996.tb01035.x

Cited by 126 publications

(114 citation statements)

References 28 publications

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“…To address this question we tested the ability of arp3-c1 cells to sporulate at the restrictive temperature of 19˚C (McCollum et al, 1996). Although arp3-c1 cells were capable of performing meiosis I and II, as evidenced from the presence of tetranucleated asci, spores were not detected in these asci (Fig.…”

Section: For3 and Arp2/3 Are Important For Meiar Assembly And Functionmentioning

confidence: 99%

Meiotic actin rings are essential for proper sporulation in fission yeast

Yan

Balasubramanian

2012

Journal of Cell Science

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SummarySporulation is a unique form of cytokinesis that occurs following meiosis II in many yeasts, during which four daughter cells (spores) are generated within a single mother cell. Here we characterize the role of F-actin in the process of sporulation in the fission yeast Schizosaccharomyces pombe. As shown previously, we find that F-actin assembles into four ring structures per ascus, referred to as the meiotic actin ring (MeiAR). The actin nucleators Arp2/3 and formin For3 assemble into ring structures that overlap with Meu14, a protein known to assemble into the so-called leading edge, a ring structure that is known to guide forespore membrane assembly. Interestingly, F-actin makes rings that occupy a larger region behind the leading edge ring. Time-lapse microscopy showed that the MeiAR assembles near the spindle pole bodies and undergoes an expansion in diameter during the early stages of meiosis II, followed by closure in later stages of meiosis II. MeiAR closure completes the process of forespore membrane assembly. Loss of the MeiAR leads to excessive assembly of forespore membranes with a deformed appearance. The rate of closure of the MeiAR is dictated by the function of the septation initiation network (SIN). We conclude that the MeiAR ensures proper targeting of the membrane biogenesis machinery to the leading edge, thereby ensuring the formation of spherical spores.

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Section: For3 and Arp2/3 Are Important For Meiar Assembly And Functionmentioning

confidence: 99%

Meiotic actin rings are essential for proper sporulation in fission yeast

Yan

Balasubramanian

2012

Journal of Cell Science

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“…Calcofluor was used to detect cell wall material in live cells as described (Balasubramanian et al, 1997). Antibodies to Arp3 (McCollum et al, 1996) and actin (Amersham) were diluted 1:100, and detected with Texas-Red-tagged anti-rabbit-IgG and Texas-Red-tagged anti-mouse-IgG antibodies (Molecular Probes) in methanol-fixed cells. DNA was visualized using DAPI as described (Balasubramanian et al, 1997).…”

Section: Fluorescence Microscopymentioning

confidence: 99%

Cell wall remodeling at the fission yeast cell division site requires the Rho-GEF Rgf3p

Morrell‐Falvey

Ren

Feoktistova

et al. 2005

Journal of Cell Science

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IntroductionCytokinesis is the final step of the cell cycle that results in the formation of two daughter cells from one. This process is regulated temporally and spatially such that the replicated and segregated chromosomes are evenly divided between daughter cells. The genetically tractable yeast Schizosaccharomyces pombe is an excellent organism for studying cytokinesis as it divides by medial fission following assembly and contraction of an actomyosin-based cleavage apparatus that directs placement and assembly of new membrane and cell wall materials (Balasubramanian et al., 2004;Krapp et al., 2004;Wolfe and Gould, 2005). Furthermore, numerous mutations are available that are defective in each step of the cell division process.The initial phase of cytokinesis involves establishing the division site at the center of a symmetrical cell. This process begins at the onset of mitosis when components of the actomyosin ring are recruited to the cell cortex immediately overlaying the nucleus (reviewed by Balasubramanian et al., 2004;Wolfe and Gould, 2005). The next steps of cytokinesis, actomyosin ring constriction and septation, require a conserved signaling cascade referred to as the septation initiation network (SIN) that is organized at the spindle pole body, a microtubuleorganizing center analogous to the mammalian centrosome (reviewed by Krapp et al., 2004;McCollum and Gould, 2001).Septation occurs concomitantly with actomyosin ring constriction as the primary septum is laid down in a centripetal manner behind the constricting CAR (reviewed by Le Goff et al., 1999a). Indeed, actomyosin ring constriction does not proceed in the absence of primary septum formation (Le Goff et al., 1999b;Liu et al., 1999). The primary septum consists primarily of unbranched 1,3-␤-glucan and 1,3-␣-glucan (Humbel et al., 2001) and Cps1p is a 1,3-␤-glucan synthase essential for its assembly (Le Goff et al., 1999b;Liu et al., 1999). , 2004). Efficient septum degradation and cell separation also require the exocyst complex, a MAPK (Pmk1p), a MAPK phosphatase (Pmp1p), calcineurin (Ppb1p), subunits of the mediator complex (Sep10p, Sep11p, and Sep15p), as well as septins and their regulator Mid2p (An et al., 2004;Hall and Russell, 2004;Sipiczki et al., 1993;Sugiura et al., 1998;Tasto et al., 2003;Toda et al., 1996;Wang et al., 2002;Yoshida et al., 1994). Precisely how these proteins are 5563Cytokinesis in Schizosaccharomyces pombe is accompanied by several stages of cell wall remodeling at the division site. Coincident with actomyosin ring constriction, primary and secondary septa are deposited and then the primary septum is degraded to release daughter cells from one another. These steps require the activities of glucan synthases and glucanases, respectively, which must be coordinated with one another to prevent cell lysis. The lad1-1 mutation undergoes cell lysis specifically at cell division owing to the absence of the Rgf3p Rho1-guanine nucleotide exchange factor (GEF) at the division site. Electron microscopic analysis indicates tha...

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“…Formins assemble unbranched filaments present both in cables that span the length of interphase cells and in the contractile ring, which constricts during cytokinesis (3,4). Arp2/3 complex nucleates branched filaments in cortical structures called actin patches, which are sites of endocytosis located at cell poles during interphase and the cleavage furrow during cytokinesis (5)(6)(7).…”

mentioning

confidence: 99%

Structure and Biochemical Properties of Fission Yeast Arp2/3 Complex Lacking the Arp2 Subunit

Nolen

Pollard

2008

Journal of Biological Chemistry

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Arp2/3 (actin-related protein 2/3) complex is a seven-subunit complex that nucleates branched actin filaments in response to cellular signals. Nucleation-promoting factors such as WASp/ Scar family proteins activate the complex by facilitating the activating conformational change and recruiting the first actin monomer for the daughter branch. Here we address the role of the Arp2 subunit in the function of Arp2/3 complex by isolating a version of the complex lacking Arp2 (Arp2⌬ Arp2/3 complex) from fission yeast. An x-ray crystal structure of the ⌬Arp2 Arp2/3 complex showed that the rest of the complex is unperturbed by the loss of Arp2. However, the Arp2⌬ Arp2/3 complex was inactive in actin nucleation assays, indicating that Arp2 is essential to form a branch. A fluorescence anisotropy assay showed that Arp2 does not contribute to the affinity of the complex for Wsp1-VCA, a Schizosaccharomyces pombe nucleationpromoting factor protein. Fluorescence resonance energy transfer experiments showed that the loss of Arp2 does not prevent VCA from recruiting an actin monomer to the complex. Truncation of the N terminus of ARPC5, the smallest subunit in the complex, increased the yield of Arp2⌬ Arp2/3 complex during purification but did not compromise nucleation activity of the full Arp2/3 complex.

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The Schizosaccharomyces pombe actin-related protein, Arp3, is a component of the cortical actin cytoskeleton and interacts with profilin.

Cited by 126 publications

References 28 publications

Meiotic actin rings are essential for proper sporulation in fission yeast

Meiotic actin rings are essential for proper sporulation in fission yeast

Cell wall remodeling at the fission yeast cell division site requires the Rho-GEF Rgf3p

Structure and Biochemical Properties of Fission Yeast Arp2/3 Complex Lacking the Arp2 Subunit

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