77 research outputs found

    Ybp2 Associates with the Central Kinetochore of Saccharomyces cerevisiae and Mediates Proper Mitotic Progression

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    The spindle checkpoint ensures the accurate segregation of chromosomes by monitoring the status of kinetochore attachment to microtubules. Simultaneous mutations in one of several kinetochore and cohesion genes and a spindle checkpoint gene cause a synthetic-lethal or synthetic-sick phenotype. A synthetic genetic array (SGA) analysis using a mad2Δ query mutant strain of yeast identified YBP2, a gene whose product shares sequence similarity with the product of YBP1, which is required for H2O2-induced oxidation of the transcription factor Yap1. ybp2Δ was sensitive to benomyl and accumulated at the mitotic stage of the cell cycle. Ybp2 physically associates with proteins of the COMA complex (Ctf19, Okp1, Mcm21, and Ame1) and 3 components of the Ndc80 complex (Ndc80, Nuf2, and Spc25 but not Spc24) in the central kinetochore and with Cse4 (the centromeric histone and CENP-A homolog). Chromatin-immunoprecipitation analyses revealed that Ybp2 associates specifically with CEN DNA. Furthermore, ybp2Δ showed synthetic-sick interactions with mutants of the genes that encode the COMA complex components. Ybp2 seems to be part of a macromolecular kinetochore complex and appears to contribute to the proper associations among the central kinetochore subcomplexes and the kinetochore-specific nucleosome

    Muon capture by 3He nuclei followed by proton and deuteron production

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    The paper describes an experiment aimed at studying muon capture by 3He{}^{3}\mathrm{He} nuclei in pure 3He{}^{3}\mathrm{He} and D2+3He\mathrm{D}_2 + {}^{3}\mathrm{He} mixtures at various densities. Energy distributions of protons and deuterons produced via μ+3Hep+n+n+νμ\mu^-+{}^{3}\mathrm{He}\to p+n+n + \nu_{\mu } and μ+3Hed+n+νμ\mu^-+{}^{3} \mathrm{He} \to d+n + \nu_{\mu} are measured for the energy intervals 104910 - 49 MeV and 133113 - 31 MeV, respectively. Muon capture rates, λcapp(ΔEp)\lambda_\mathrm{cap}^p (\Delta E_p) and λcapd(ΔEd)\lambda_\mathrm{cap}^d (\Delta E_d) are obtained using two different analysis methods. The least--squares methods gives λcapp=(36.7±1.2)s1\lambda_\mathrm{cap}^p = (36.7\pm 1.2) {s}^{- 1}, λcapd=(21.3±1.6)s1\lambda_\mathrm{cap}^d = (21.3 \pm 1.6) {s}^{- 1}. The Bayes theorem gives λcapp=(36.8±0.8)s1\lambda_\mathrm{cap}^p = (36.8 \pm 0.8) {s}^{- 1}, λcapd=(21.9±0.6)s1\lambda_\mathrm{cap}^d = (21.9 \pm 0.6) {s}^{- 1}. The experimental differential capture rates, dλcapp(Ep)/dEpd\lambda_\mathrm{cap}^p (E_p) / dE_p and dλcapd(Ed)/dEd d\lambda_\mathrm{cap}^d (E_d) / dE_d, are compared with theoretical calculations performed using the plane--wave impulse approximation (PWIA) with the realistic NN interaction Bonn B potential. Extrapolation to the full energy range yields total proton and deuteron capture rates in good agreement with former results.Comment: 17 pages, 13 figures, accepted for publication in PR

    Quasi-Elastic Scattering in the Inclusive (3^3He, t) Reaction

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    The triton energy spectra of the charge-exchange 12^{12}C(3^3He,t) reaction at 2 GeV beam energy are analyzed in the quasi-elastic nucleon knock-out region. Considering that this region is mainly populated by the charge-exchange of a proton in 3^3He with a neutron in the target nucleus and the final proton going in the continuum, the cross-sections are written in the distorted-wave impulse approximation. The t-matrix for the elementary exchange process is constructed in the DWBA, using one pion- plus rho-exchange potential for the spin-isospin nucleon- nucleon potential. This t-matrix reproduces the experimental data on the elementary pn \rightarrow np process. The calculated cross-sections for the 12^{12}C(3^3He,t) reaction at 2o2^o to 7o7^o triton emission angle are compared with the corresponding experimental data, and are found in reasonable overall accord.Comment: 19 pages, latex, 11 postscript figures available at [email protected], submitted to Phy.Rev.

    Induced pseudoscalar coupling of the proton weak interaction

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    The induced pseudoscalar coupling gpg_p is the least well known of the weak coupling constants of the proton's charged--current interaction. Its size is dictated by chiral symmetry arguments, and its measurement represents an important test of quantum chromodynamics at low energies. During the past decade a large body of new data relevant to the coupling gpg_p has been accumulated. This data includes measurements of radiative and non radiative muon capture on targets ranging from hydrogen and few--nucleon systems to complex nuclei. Herein the authors review the theoretical underpinnings of gpg_p, the experimental studies of gpg_p, and the procedures and uncertainties in extracting the coupling from data. Current puzzles are highlighted and future opportunities are discussed.Comment: 58 pages, Latex, Revtex4, prepared for Reviews of Modern Physic

    The Rts1 Regulatory Subunit of Protein Phosphatase 2A Is Required for Control of G1 Cyclin Transcription and Nutrient Modulation of Cell Size

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    The key molecular event that marks entry into the cell cycle is transcription of G1 cyclins, which bind and activate cyclin-dependent kinases. In yeast cells, initiation of G1 cyclin transcription is linked to achievement of a critical cell size, which contributes to cell-size homeostasis. The critical cell size is modulated by nutrients, such that cells growing in poor nutrients are smaller than cells growing in rich nutrients. Nutrient modulation of cell size does not work through known critical regulators of G1 cyclin transcription and is therefore thought to work through a distinct pathway. Here, we report that Rts1, a highly conserved regulatory subunit of protein phosphatase 2A (PP2A), is required for normal control of G1 cyclin transcription. Loss of Rts1 caused delayed initiation of bud growth and delayed and reduced accumulation of G1 cyclins. Expression of the G1 cyclin CLN2 from an inducible promoter rescued the delayed bud growth in rts1Δ cells, indicating that Rts1 acts at the level of transcription. Moreover, loss of Rts1 caused altered regulation of Swi6, a key component of the SBF transcription factor that controls G1 cyclin transcription. Epistasis analysis revealed that Rts1 does not work solely through several known critical upstream regulators of G1 cyclin transcription. Cells lacking Rts1 failed to undergo nutrient modulation of cell size. Together, these observations demonstrate that Rts1 is a key player in pathways that link nutrient availability, cell size, and G1 cyclin transcription. Since Rts1 is highly conserved, it may function in similar pathways in vertebrates

    Mutability and mutational spectrum of chromosome transmission fidelity genes

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    It has been more than two decades since the original chromosome transmission fidelity (Ctf) screen of Saccharomyces cerevisiae was published. Since that time the spectrum of mutations known to cause Ctf and, more generally, chromosome instability (CIN) has expanded dramatically as a result of systematic screens across yeast mutant arrays. Here we describe a comprehensive summary of the original Ctf genetic screen and the cloning of the remaining complementation groups as efforts to expand our knowledge of the CIN gene repertoire and its mutability in a model eukaryote. At the time of the original screen, it was impossible to predict either the genes and processes that would be overrepresented in a pool of random mutants displaying a Ctf phenotype or what the entire set of genes potentially mutable to Ctf would be. We show that in a collection of 136 randomly selected Ctf mutants, >65% of mutants map to 13 genes, 12 of which are involved in sister chromatid cohesion and/or kinetochore function. Extensive screening of systematic mutant collections has shown that ~350 genes with functions as diverse as RNA processing and proteasomal activity mutate to cause a Ctf phenotype and at least 692 genes are required for faithful chromosome segregation. The enrichment of random Ctf alleles in only 13 of ~350 possible Ctf genes suggests that these genes are more easily mutable to cause genome instability than the others. These observations inform our understanding of recurring CIN mutations in human cancers where presumably random mutations are responsible for initiating the frequently observed CIN phenotype of tumors

    Theoretical Study of the ^3He(mu^-,nu_mu)^3H Capture

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    The ^3He(mu^-,nu_mu)^3H weak capture is studied using correlated-hyperspherical-harmonics wave functions, obtained from realistic Hamiltonians consisting of the Argonne v14v_{14} or Argonne v18v_{18} two-nucleon, and Tucson-Melbourne or Urbana-IX three-nucleon interactions. The nuclear weak charge and current operators have vector and axial-vector components that include one- and two-body contributions. The strength of the leading two-body operator in the axial-vector current is adjusted to reproduce the Gamow-Teller matrix element in tritium β\beta-decay. The calculated total capture rate is in excellent agreement with the most recent experimental determination 1496±41496\pm 4 sec1^{-1}, when the PCAC value is adopted for the induced pseudo-scalar coupling constant gPSg_{PS}. The predictions for the capture rate and angular correlation parameters AvA_v, AtA_t, and AΔA_\Delta are found to be only very weakly dependent on the model input Hamiltonian. The variation of these observables with gPSg_{PS} and the theoretical uncertainties deriving from the model-dependent procedure used to constrain the axial current are investigated.Comment: 16 pages, 1 figure, submitted to PR

    Roles for the Conserved Spc105p/Kre28p Complex in Kinetochore-Microtubule Binding and the Spindle Assembly Checkpoint

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    Kinetochores attach sister chromatids to microtubules of the mitotic spindle and orchestrate chromosome disjunction at anaphase. Although S. cerevisiae has the simplest known kinetochores, they nonetheless contain approximately 70 subunits that assemble on centromeric DNA in a hierarchical manner. Developing an accurate picture of the DNA-binding, linker and microtubule-binding layers of kinetochores, including the functions of individual proteins in these layers, is a key challenge in the field of yeast chromosome segregation. Moreover, comparison of orthologous proteins in yeast and humans promises to extend insight obtained from the study of simple fungal kinetochores to complex animal cell kinetochores.We show that S. cerevisiae Spc105p forms a heterotrimeric complex with Kre28p, the likely orthologue of the metazoan kinetochore protein Zwint-1. Through systematic analysis of interdependencies among kinetochore complexes, focused on Spc105p/Kre28p, we develop a comprehensive picture of the assembly hierarchy of budding yeast kinetochores. We find Spc105p/Kre28p to comprise the third linker complex that, along with the Ndc80 and MIND linker complexes, is responsible for bridging between centromeric heterochromatin and kinetochore MAPs and motors. Like the Ndc80 complex, Spc105p/Kre28p is also essential for kinetochore binding by components of the spindle assembly checkpoint. Moreover, these functions are conserved in human cells.Spc105p/Kre28p is the last of the core linker complexes to be analyzed in yeast and we show it to be required for kinetochore binding by a discrete subset of kMAPs (Bim1p, Bik1p, Slk19p) and motors (Cin8p, Kar3p), all of which are nonessential. Strikingly, dissociation of these proteins from kinetochores prevents bipolar attachment, even though the Ndc80 and DASH complexes, the two best-studied kMAPs, are still present. The failure of Spc105 deficient kinetochores to bind correctly to spindle microtubules and to recruit checkpoint proteins in yeast and human cells explains the observed severity of missegregation phenotypes
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