Cdc4p, a contractile ring protein essential for cytokinesis in Schizosaccharomyces pombe, interacts with a phosphatidylinositol 4-kinase.

The proposed function of Cdc4p, an essential contractile ring protein in Schizosaccharomyces pombe, is that of a myosin essential light chain. However, five conditionally lethal cdc4 alleles exhibit complementation in diploids. Such interallelic complementation is not readily explained if the sole function of Cdc4p is that of a myosin essential light chain. Complementation of cdc4 alleles could occur only if different mutant forms can assemble into an active oligomeric complex or if Cdc4p has more than one essential function. To search for other proteins that may interact with Cdc4p, we performed a two-hybrid screen and identified two such candidates: one similar to Saccharomyces cerevisiae Vps27p and the other a putative phosphatidylinositol (PI) 4-kinase. Binding of Cdc4p to the latter and to myosin heavy chain (Myo2p) was confirmed by immunosorbent assays. Deletion studies demonstrated interaction between the Cdc4p C-terminal domain and the PI 4-kinase C-terminal domain. Furthermore, interaction was abolished by the Cdc4p C-terminal domain point mutation, Gly107 to Ser. This allele also causes failure of cytokinesis. Ectopic expression of the PI 4-kinase C-terminal domain caused cytokinesis defects that were most extreme in cells carrying the G107S allele. We suggest that Cdc4p plays multiple roles in cytokinesis and that interaction with a PI 4-kinase may be important for contractile ring assembly and/or function

Structure of Cdc4p, a Contractile Ring Protein Essential for Cytokinesis in Schizosaccharomyces pombe

The Schizosaccharomyces pombe Cdc4 protein is required for the formation and function of the contractile ring, presumably acting as a myosin light chain. By using NMR spectroscopy, we demonstrate that purified Cdc4p is a monomeric protein with two structurally independent domains, each exhibiting a fold reminiscent of the EF-hand class of calcium-binding proteins. Although Cdc4p has one potentially functional calcium-binding site, it does not bind calcium in vitro. Three variants of Cdc4p containing single point mutations responsible for temperature-sensitive arrest of the cell cycle at cytokinesis (Gly-19 to Glu, Gly-82 to Asp, and Gly-107 to Ser) were also characterized by NMR and circular dichroism spectroscopy. In each case, the amino acid substitution only leads to small perturbations in the conformation of the protein. Furthermore, thermal unfolding studies indicate that, like wild-type Cdc4p, the three mutant forms are all extremely stable, remaining completely folded at temperatures significantly above those causing failure of cytokinesis in intact cells. Therefore, the altered phenotype must arise directly from a disruption of the function of Cdc4p rather than indirectly through a disruption of its overall structure. Several mutant alleles of Cdc4p also show interallelic complementation in diploid cells. This phenomenon can be explained if Cdcp4 has more than one essential function or, alternatively, if two mutant proteins assemble to form a functional complex. Based on the structure of Cdc4p, possible models for interallelic complementation including interactions with partner proteins and the formation of a myosin complex with Cdc4p fulfilling the role of both an essential and regulatory light chain are proposed

Observed Effect of Magnetic Fields on the Propagation of Magnetoacoustic Waves in the Lower Solar Atmosphere

We study Hinode/SOT-FG observations of intensity fluctuations in Ca II H-line and G-band image sequences and their relation to simultaneous and co-spatial magnetic field measurements. We explore the G-band and H-line intensity oscillation spectra both separately and comparatively via their relative phase differences, time delays and cross-coherences. In the non-magnetic situations, both sets of fluctuations show strong oscillatory power in the 3 - 7 mHz band centered at 4.5 mHz, but this is suppressed as magnetic field increases. A relative phase analysis gives a time delay of H-line after G-band of 20\pm1 s in non-magnetic situations implying a mean effective height difference of 140 km. The maximum coherence is at 4 - 7 mHz. Under strong magnetic influence the measured delay time shrinks to 11 s with the peak coherence near 4 mHz. A second coherence maximum appears between 7.5 - 10 mHz. Investigation of the locations of this doubled-frequency coherence locates it in diffuse rings outside photospheric magnetic structures. Some possible interpretations of these results are offered.Comment: 19 pages, 6 figure

O-GlcNAc Modification of tau Directly Inhibits Its Aggregation without Perturbing the Conformational Properties of tau Monomers

Abstract The aggregation of the microtubule-associated protein tau into paired helical filaments to form neurofibrillary tangles constitutes one of the pathological hallmarks of Alzheimer's disease. Tau is post-translationally modified by the addition of N-acetyl-D-glucosamine O-linked to several serine and threonine residues (O-GlcNAc). Previously, increased O-GlcNAcylation of tau has been shown to block the accumulation of tau aggregates within a tauopathy mouse model. Here we show that O-GlcNAc modification of full-length human tau impairs the rate and extent of its heparin-induced aggregation without perturbing its activity toward microtubule polymerization. O-GlcNAcylation, however, does not impact the "global-fold" of tau as measured by a Förster resonance energy transfer assay. Similarly, nuclear magnetic resonance studies demonstrated that O-GlcNAcylation only minimally perturbs the local structural and dynamic features of a tau fragment (residues 353-408) spanning the last microtubule binding repeat to the major GlcNAc-acceptor Ser400. These data indicate that the inhibitory effects of O-GlcNAc on tau aggregation may result from enhanced monomer solubility or the destabilization of fibrils or soluble aggregates, rather than by altering the conformational properties of the monomeric protein. This work further underscores the potential of targeting the O-GlcNAc pathway for potential Alzheimer's disease therapeutics

O-GlcNAc Modification of tau Directly Inhibits Its Aggregation without Perturbing the Conformational Properties of tau Monomers

The aggregation of the microtubule-associated protein tau into paired helical filaments to form neurofibrillary tangles constitutes one of the pathological hallmarks of Alzheimer\u27s disease. Tau is post-translationally modified by the addition of N-acetyl-d-glucosamine O-linked to several serine and threonine residues (O-GlcNAc). Previously, increased O-GlcNAcylation of tau has been shown to block the accumulation of tau aggregates within a tauopathy mouse model. Here we show that O-GlcNAc modification of full-length human tau impairs the rate and extent of its heparin-induced aggregation without perturbing its activity toward microtubule polymerization. O-GlcNAcylation, however, does not impact the “global-fold” of tau as measured by a Förster resonance energy transfer assay. Similarly, nuclear magnetic resonance studies demonstrated that O-GlcNAcylation only minimally perturbs the local structural and dynamic features of a tau fragment (residues 353–408) spanning the last microtubule binding repeat to the major GlcNAc-acceptor Ser400. These data indicate that the inhibitory effects of O-GlcNAc on tau aggregation may result from enhanced monomer solubility or the destabilization of fibrils or soluble aggregates, rather than by altering the conformational properties of the monomeric protein. This work further underscores the potential of targeting the O-GlcNAc pathway for potential Alzheimer\u27s disease therapeutics

Multiresolution analysis of active region magnetic structure and its correlation with the Mt. Wilson classification and flaring activity

Two different multi-resolution analyses are used to decompose the structure of active region magnetic flux into concentrations of different size scales. Lines separating these opposite polarity regions of flux at each size scale are found. These lines are used as a mask on a map of the magnetic field gradient to sample the local gradient between opposite polarity regions of given scale sizes. It is shown that the maximum, average and standard deviation of the magnetic flux gradient for alpha, beta, beta-gamma and beta-gamma-delta active regions increase in the order listed, and that the order is maintained over all length-scales. This study demonstrates that, on average, the Mt. Wilson classification encodes the notion of activity over all length-scales in the active region, and not just those length-scales at which the strongest flux gradients are found. Further, it is also shown that the average gradients in the field, and the average length-scale at which they occur, also increase in the same order. Finally, there are significant differences in the gradient distribution, between flaring and non-flaring active regions, which are maintained over all length-scales. It is also shown that the average gradient content of active regions that have large flares (GOES class 'M' and above) is larger than that for active regions containing flares of all flare sizes; this difference is also maintained at all length-scales.Comment: Accepted for publication in Solar Physic

Turbulence in the Solar Atmosphere: Manifestations and Diagnostics via Solar Image Processing

Intermittent magnetohydrodynamical turbulence is most likely at work in the magnetized solar atmosphere. As a result, an array of scaling and multi-scaling image-processing techniques can be used to measure the expected self-organization of solar magnetic fields. While these techniques advance our understanding of the physical system at work, it is unclear whether they can be used to predict solar eruptions, thus obtaining a practical significance for space weather. We address part of this problem by focusing on solar active regions and by investigating the usefulness of scaling and multi-scaling image-processing techniques in solar flare prediction. Since solar flares exhibit spatial and temporal intermittency, we suggest that they are the products of instabilities subject to a critical threshold in a turbulent magnetic configuration. The identification of this threshold in scaling and multi-scaling spectra would then contribute meaningfully to the prediction of solar flares. We find that the fractal dimension of solar magnetic fields and their multi-fractal spectrum of generalized correlation dimensions do not have significant predictive ability. The respective multi-fractal structure functions and their inertial-range scaling exponents, however, probably provide some statistical distinguishing features between flaring and non-flaring active regions. More importantly, the temporal evolution of the above scaling exponents in flaring active regions probably shows a distinct behavior starting a few hours prior to a flare and therefore this temporal behavior may be practically useful in flare prediction. The results of this study need to be validated by more comprehensive works over a large number of solar active regions.Comment: 26 pages, 7 figure

Autoinhibition of ETV6 (TEL) DNA binding: appended helices sterically block the ETS domain.

ETV6 (or TEL), a transcriptional repressor belonging to the ETS family, is frequently involved in chromosomal translocations linked with human cancers. It displays a DNA-binding mode distinct from other ETS proteins due to the presence of a self-associating PNT domain. In this study, we used NMR spectroscopy to dissect the structural and dynamic bases for the autoinhibition of ETV6 DNA binding by sequences C-terminal to its ETS domain. The C-terminal inhibitory domain (CID) contains two helices, H4 and H5, which sterically block the DNA-binding interface of the ETS domain. Importantly, these appended helices are only marginally stable as revealed by amide hydrogen exchange and 15 N relaxation measurements. The CID is thus poised to undergo a facile conformational change as required for DNA binding. The CID also dampens millisecond timescale motions of the ETS domain hypothesized to be critical for the recognition of specific ETS target sequences. This work illustrates the use of appended sequences on conserved structural domains to generate biological diversity and complements previous studies of the allosteric mechanism of ETS1 autoinhibition to reveal both common and divergent features underlying the regulation of DNA binding by ETS transcription factors

The solar chromosphere at high resolution with IBIS. II. Acoustic shocks in the quiet internetwork and the role of magnetic fields

(Abridged) Aims: We characterize the dynamics of the quiet inter-network chromosphere by studying the occurrence of acoustic shocks and their relation with the concomitant photospheric structure and dynamics. Methods: We analyze a comprehensive data set that includes high resolution chromospheric and photospheric spectra obtained with the IBIS imaging spectrometer in two quiet-Sun regions. This is complemented by high-resolution sequences of MDI magnetograms of the same targets. From the chromospheric spectra we identify the spatio-temporal occurrence of the acoustic shocks. We compare it with the photospheric dynamics by means of both Fourier and wavelet analysis, and study the influence of magnetic structures. Results: Mid-chromospheric shocks occur as a response to underlying powerful photospheric motions at periodicities nearing the acoustic cut-off, consistent with 1-D hydrodynamical modeling. However, their spatial distribution within the supergranular cells is highly dependent on the local magnetic topology, both at the network and internetwork scale. Large portions of the internetwork regions undergo very few shocks, as "shadowed" by the horizontal component of the magnetic field. The latter is betrayed by the presence of chromospheric fibrils, observed in the core of the CaII line as slanted structures with distinct dynamical properties. The shadow mechanism appears to operate also on the very small scales of inter-network magnetic elements, and provides for a very pervasive influence of the magnetic field even in the quietest region analyzed.Comment: 18 pages, 16 figures (includes 1 Appendix). Accepted by A&A (16 October 2008). Minor changes from v1 after referee's comments. Higher quality figures available at http://www.arcetri.astro.it/~gcauzzi/papers_astroph/ibis.shocks.accepted.pd