Probing the environment of emerin by Enhanced ascorbate peroxidase 2 (APEX2)-mediated proximity labeling.

Emerin is one of the best characterized proteins of the inner nuclear membrane, but can also occur at the level of the endoplasmic reticulum. We now use enhanced ascorbate peroxidase 2 (APEX2) to probe the environment of emerin. APEX2 can be used as a genetic tag that produces short-lived yet highly reactive biotin species, allowing the modification of proteins that interact with or are in very close proximity to the tagged protein. Biotinylated proteins can be isolated using immobilized streptavidin and analyzed by mass spectrometry. As an alternative to the standard approach with a genetic fusion of APEX2 to emerin, we also used RAPIDS (rapamycin- and APEX-dependent identification of proteins by SILAC), a method with improved specificity, where the peroxidase interacts with the protein of interest (i.e., emerin) only upon addition of rapamycin to the cells. We compare these different approaches, which, together, identify well-known interaction partners of emerin like lamin A and the lamina associated polypeptide 1 (LAP1), as well as novel proximity partners

Effects of temporal variability of disturbance on the succession in marine fouling communities in northern-central Chile

We investigated the effects of temporal variability in a disturbance regime on fouling communities at two study sites in a northern-central Chilean bay. Fouling assemblages grown on artificial settlement substrata were disturbed by mechanical removal of biomass at different time intervals. Using one single disturbance frequency (10 disturbance events over 5 months) we applied 7 different temporal disturbance treatments: a constant disturbance regime (identical intervals between disturbance events), and 6 variable treatments where both variableness and sequences of intervals between disturbance events were manipulated. Two levels of temporal variableness (low and high, i.e. disturbance events were either dispersed or highly clumped in time) in the disturbance regime were applied by modifying the time intervals between subsequent disturbance events. To investigate the temporal coupling between disturbance events and other ecological processes (e.g. larval supply and recruitment intensity), three different sequences of disturbance intervals were nested in each of the two levels of temporal variableness. Species richness, evenness, total abundance, and structure of communities that experienced the various disturbance regimes were compared at the end of the experiment (15 days after the last disturbance event). Disturbance strongly influenced the community structure and led to a decrease in evenness and total abundance but not species richness. In undisturbed reference communities, the dominant competitor Pyura chilensis (Tunicata) occupied most available space while this species was suppressed in all disturbed treatments. Surprisingly, neither temporal variableness in the disturbance regime nor the sequence of intervals between disturbance events had an effect on community structure. Temporal variability in high disturbance regimes may be of minor importance for fouling communities, because they are dominated by opportunistic species that are adapted to rapidly exploit available space

Extinctions and Correlations for Uniformly Discrete Point Processes with Pure Point Dynamical Spectra

The paper investigates how correlations can completely specify a uniformly discrete point process. The setting is that of uniformly discrete point sets in real space for which the corresponding dynamical hull is ergodic. The first result is that all of the essential physical information in such a system is derivable from its $n$-point correlations, $n= 2, 3, >...$. If the system is pure point diffractive an upper bound on the number of correlations required can be derived from the cycle structure of a graph formed from the dynamical and Bragg spectra. In particular, if the diffraction has no extinctions, then the 2 and 3 point correlations contain all the relevant information.Comment: 16 page

Quantum Electrodynamics in the Light-Front Weyl Gauge

We examine QED(3+1) quantised in the `front form' with finite `volume' regularisation, namely in Discretised Light-Cone Quantisation. Instead of the light-cone or Coulomb gauges, we impose the light-front Weyl gauge $A^-=0$. The Dirac method is used to arrive at the quantum commutation relations for the independent variables. We apply `quantum mechanical gauge fixing' to implement Gau{\ss}' law, and derive the physical Hamiltonian in terms of unconstrained variables. As in the instant form, this Hamiltonian is invariant under global residual gauge transformations, namely displacements. On the light-cone the symmetry manifests itself quite differently.Comment: LaTeX file, 30 pages (A4 size), no figures. Submitted to Physical review D. January 18, 1996. Originally posted, erroneously, with missing `Weyl' in title. Otherwise, paper is identica

Cloning and characterisation of chlorophyll synthase from Avena sativa

The chlorophyll synthase gene from oat (Avena sativa) was cloned and expressed in Escherichia coli. The deduced amino acid sequence consists of 378 amino acids including a presequence, of 46 amino acids. Deletion mutants show that a core protein comprising amino acid residues 88 to 377 is enzymatically active. The sequence of the mature protein shows 85% identity with the chlorophyll synthase of Arabidopsis thaliana and 62% identity with the chlorophyll synthase of Synechocystis PCC 6803. The gene is constitutively expressed as the same transcript level is found in dark-grown and in light-grown seedlings. The enzyme requires magnesium ions for activity; manganese ions can reconstitute only part of the activity. Diacetyl and N-phenylmaleimide (NPM) inhibit the enzyme activity. Site-directed mutagenesis reveals that, out of the 4 Arg residues present in the active core protein, Arg-91 and Arg-161 are essential for the activity. Five cysteine residues are present in the core protein, of which only Cys-109 is essential for the enzyme activity. Since the wild-type and all other Cys-mutants with the exception of the mutant C304A are inhibited by N-phenylmaleimide, we conclude that the inhibitor binds to a non-essential Cys residue to abolish activity. The role of the various Arg and Cys residues is discussed

The Path Integral for 1+1-dimensional QCD

We derive a path integral expression for the transition amplitude in 1+1-dimensional QCD starting from canonically quantized QCD. Gauge fixing after quantization leads to a formulation in terms of gauge invariant but curvilinear variables. Remainders of the curved space are Jacobians, an effective potential, and sign factors just as for the problem of a particle in a box. Based on this result we derive a Faddeev-Popov like expression for the transition amplitude avoiding standard infinities that are caused by integrations over gauge equivalent configurations.Comment: 16 pages, LaTeX, 3 PostScript figures, uses epsf.st

A cross impact methodology for the assessment of US telecommunications system with application to fiber optics development: Executive summary

A cross impact model of the U.S. telecommunications system was developed. For this model, it was necessary to prepare forecasts of the major segments of the telecommunications system, such as satellites, telephone, TV, CATV, radio broadcasting, etc. In addition, forecasts were prepared of the traffic generated by a variety of new or expanded services, such as electronic check clearing and point of sale electronic funds transfer. Finally, the interactions among the forecasts were estimated (the cross impacts). Both the forecasts and the cross impacts were used as inputs to the cross impact model, which could then be used to stimulate the future growth of the entire U.S. telecommunications system. By varying the inputs, technology changes or policy decisions with regard to any segment of the system could be evaluated in the context of the remainder of the system. To illustrate the operation of the model, a specific study was made of the deployment of fiber optics, throughout the telecommunications system

QCD near the Light Cone

Starting from the QCD Lagrangian, we present the QCD Hamiltonian for near light cone coordinates. We study the dynamics of the gluonic zero modes of this Hamiltonian. The strong coupling solutions serve as a basis for the complete problem. We discuss the importance of zero modes for the confinement mechanism.Comment: 32 pages, ReVTeX, 2 Encapsulated PostScript figure