5,205 research outputs found

    Analysis of Mitotic Microtubule-Associated Proteins Using Mass Spectrometry Identifies Astrin, a Spindle-Associated Protein

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    We purified microtubules from a mammalian mitotic extract and obtained an amino acid sequence from each microtubule-associated protein by using mass spectrometry. Most of these proteins are known spindle-associated components with essential functional roles in spindle organization. We generated antibodies against a protein identified in this collection and refer to it as astrin because of its association with astral microtubule arrays assembled in vitro. Astrin is approximately 134 kDa, and except for a large predicted coiled-coil domain in its C-terminal region it lacks any known functional motifs. Astrin associates with spindle microtubules as early as prophase where it concentrates at spindle poles. It localizes throughout the spindle in metaphase and anaphase and associates with midzone microtubules in anaphase and telophase. Astrin also localizes to kinetochores but only on those chromosomes that have congressed. Deletion analysis indicates that astrin\u27s primary spindle-targeting domain is at the C terminus, although a secondary domain in the N terminus can target some of the protein to spindle poles. Thus, we have generated a comprehensive list of major mitotic microtubule-associated proteins, among which is astrin, a nonmotor spindle protein

    The KinI kinesin Kif2a is required for bipolar spindle assembly through a functional relationship with MCAK

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    Although the microtubule-depolymerizing KinI motor Kif2a is abundantly expressed in neuronal cells, we now show it localizes to centrosomes and spindle poles during mitosis in cultured cells. RNAi-induced knockdown of Kif2a expression inhibited cell cycle progression because cells assembled monopolar spindles. Bipolar spindle assembly was restored in cells lacking Kif2a by treatments that altered microtubule assembly (nocodazole), eliminated kinetochore–microtubule attachment (loss of Nuf2), or stabilized microtubule plus ends at kinetochores (loss of MCAK). Thus, two KinI motors, MCAK and Kif2a, play distinct roles in mitosis, and MCAK activity at kinetochores must be balanced by Kif2a activity at poles for spindle bipolarity. These treatments failed to restore bipolarity to cells lacking the activity of the kinesin Eg5. Thus, two independent pathways contribute to spindle bipolarity, with the Eg5-dependent pathway using motor force to drive spindle bipolarity and the Kif2a-dependent pathway relying on microtubule polymer dynamics to generate force for spindle bipolarity

    A Bose-Einstein Condensate in a Uniform Light-induced Vector Potential

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    We use a two-photon dressing field to create an effective vector gauge potential for Bose-condensed Rb atoms in the F=1 hyperfine ground state. The dressed states in this Raman field are spin and momentum superpositions, and we adiabatically load the atoms into the lowest energy dressed state. The effective Hamiltonian of these neutral atoms is like that of charged particles in a uniform magnetic vector potential, whose magnitude is set by the strength and detuning of Raman coupling. The spin and momentum decomposition of the dressed states reveals the strength of the effective vector potential, and our measurements agree quantitatively with a simple single-particle model. While the uniform effective vector potential described here corresponds to zero magnetic field, our technique can be extended to non-uniform vector potentials, giving non-zero effective magnetic fields.Comment: 5 pages, submitted to Physical Review Letter

    Quantitative localized proton-promoted dissolution kinetics of calcite using scanning electrochemical microscopy (SECM)

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    Scanning electrochemical microscopy (SECM) has been used to determine quantitatively the kinetics of proton-promoted dissolution of the calcite (101̅4) cleavage surface (from natural “Iceland Spar”) at the microscopic scale. By working under conditions where the probe size is much less than the characteristic dislocation spacing (as revealed from etching), it has been possible to measure kinetics mainly in regions of the surface which are free from dislocations, for the first time. To clearly reveal the locations of measurements, studies focused on cleaved “mirror” surfaces, where one of the two faces produced by cleavage was etched freely to reveal defects intersecting the surface, while the other (mirror) face was etched locally (and quantitatively) using SECM to generate high proton fluxes with a 25 μm diameter Pt disk ultramicroelectrode (UME) positioned at a defined (known) distance from a crystal surface. The etch pits formed at various etch times were measured using white light interferometry to ascertain pit dimensions. To determine quantitative dissolution kinetics, a moving boundary finite element model was formulated in which experimental time-dependent pit expansion data formed the input for simulations, from which solution and interfacial concentrations of key chemical species, and interfacial fluxes, could then be determined and visualized. This novel analysis allowed the rate constant for proton attack on calcite, and the order of the reaction with respect to the interfacial proton concentration, to be determined unambiguously. The process was found to be first order in terms of interfacial proton concentration with a rate constant k = 6.3 (± 1.3) × 10–4 m s–1. Significantly, this value is similar to previous macroscopic rate measurements of calcite dissolution which averaged over large areas and many dislocation sites, and where such sites provided a continuous source of steps for dissolution. Since the local measurements reported herein are mainly made in regions without dislocations, this study demonstrates that dislocations and steps that arise from such sites are not needed for fast proton-promoted calcite dissolution. Other sites, such as point defects, which are naturally abundant in calcite, are likely to be key reaction sites

    Effect of Fish Size on Prey Size Selection in Gambusia Affinis

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    Food size selection of the mosquitofish , Gambusia affinis affinis. was measured in aquaria using juvenile stages of the mosquito, Clllex tarsalb;, as prey. Fish size varied from recently born fry to large adult females. Food size selection was positively correlated with fish size. Mosquitofish fry (6-8 111m standard length) attacked and ate primarily first and second instar larvae. Fry attacked larger instars, but attack success on these was low (0 - 50%). Fish larger than 20 mm attached primarily pupae and third and fourth instar larva. No first instar mosquitoes were eaten. Attack success for these fish was above 65\u27Yr) for all instars
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