The Chlamydia trachomatis Type III Secretion Chaperone Slc1 Engages Multiple Early Effectors, Including TepP, a Tyrosine-phosphorylated Protein Required for the Recruitment of CrkI-II to Nascent Inclusions and Innate Immune Signaling

Chlamydia trachomatis, the causative agent of trachoma and sexually transmitted infections, employs a type III secretion (T3S) system to deliver effector proteins into host epithelial cells to establish a replicative vacuole. Aside from the phosphoprotein TARP, a Chlamydia effector that promotes actin re-arrangements, very few factors mediating bacterial entry and early inclusion establishment have been characterized. Like many T3S effectors, TARP requires a chaperone (Slc1) for efficient translocation into host cells. In this study, we defined proteins that associate with Slc1 in invasive C. trachomatis elementary bodies (EB) by immunoprecipitation coupled with mass spectrometry. We identified Ct875, a new Slc1 client protein and T3S effector, which we renamed TepP (Translocated early phosphoprotein). We provide evidence that T3S effectors form large molecular weight complexes with Scl1 in vitro and that Slc1 enhances their T3S-dependent secretion in a heterologous Yersinia T3S system. We demonstrate that TepP is translocated early during bacterial entry into epithelial cells and is phosphorylated at tyrosine residues by host kinases. However, TepP phosphorylation occurs later than TARP, which together with the finding that Slc1 preferentially engages TARP in EBs leads us to postulate that these effectors are translocated into the host cell at different stages during C.trachomatis invasion. TepP co-immunoprecipitated with the scaffolding proteins CrkI-II during infection and Crk was recruited to EBs at entry sites where it remained associated with nascent inclusions. Importantly, C. trachomatis mutants lacking TepP failed to recruit CrkI-II to inclusions, providing genetic confirmation of a direct role for this effector in the recruitment of a host factor. Finally, endocervical epithelial cells infected with a tepP mutant showed altered expression of a subset of genes associated with innate immune responses. We propose a model wherein TepP acts downstream of TARP to recruit scaffolding proteins at entry sites to initiate and amplify signaling cascades important for the regulation of innate immune responses to Chlamydia.Fil: Chen, Yi-Shan. University of Duke; Estados UnidosFil: Bastidas, Robert J.. University of Duke; Estados UnidosFil: Saka, Hector Alex. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. University of Duke; Estados UnidosFil: Carpenter, Victoria K.. Duke University Medical Center; . University of Duke; Estados UnidosFil: Richards, Kristian L.. Miami University; Estados UnidosFil: Plano, Gregory V.. Miami University; Estados UnidosFil: Valdivia, Raphael H.. University of Duke; Estados Unido

The Influence of Marketing Mix, Perceived Risk, and Satisfaction on Word of Mouth in Xyz Clinic

The increasing need for health services, peoples who lived in the Pekayon, Bekasi City were given the opportunity to choose the right clinic. Word of mouth is a marketing technique that can be used by clinics. This study aims to analyze the effects of the marketing mix, perceived risk, and satisfaction on word of mouth at XYZ clinic. The research is a descriptive method with a survey using questionnaires and 200 respondents as the sample. Furthermore, the data analysis technique is descriptive with SPSS16.0 software and Structural Equation Model (SEM) with LISREL 8.70. Based on the results, it can be concluded that the marketing mix has a positive effect on perceived risk, marketing mix has a positive effect on satisfaction, perceived risk has a negative effect on satisfaction, marketing mix has a positive effect on word of mouth, perceived risk has a negative effect on word of mouth, and satisfaction has a positive effect on word of mouth. Referring to these conclusions, it can be confirmed that the clinical management of doctor XYZ needs to improve employee services, convenience the patient that this clinic has expert doctors, and utilizing the use of social media as a marketing strategy

Accurate structure factors from pseudopotential methods

Highly accurate experimental structure factors of silicon are available in the literature, and these provide the ideal test for any \emph{ab initio} method for the construction of the all-electron charge density. In a recent paper [J. R. Trail and D. M. Bird, Phys. Rev. B {\bf 60}, 7863 (1999)] a method has been developed for obtaining an accurate all-electron charge density from a first principles pseudopotential calculation by reconstructing the core region of an atom of choice. Here this method is applied to bulk silicon, and structure factors are derived and compared with experimental and Full-potential Linear Augmented Plane Wave results (FLAPW). We also compare with the result of assuming the core region is spherically symmetric, and with the result of constructing a charge density from the pseudo-valence density + frozen core electrons. Neither of these approximations provide accurate charge densities. The aspherical reconstruction is found to be as accurate as FLAPW results, and reproduces the residual error between the FLAPW and experimental results.Comment: 6 Pages, 3 figure

N=2 Instanton Effective Action in Omega-background and D3/D(-1)-brane System in R-R Background

We study the relation between the ADHM construction of instantons in the Omega-background and the fractional D3/D(-1)-branes at the orbifold singularity of C \times C^2/Z_2 in Ramond-Ramond (R-R) 3-form field strength background. We calculate disk amplitudes of open strings connecting the D3/D(-1)-branes in certain R-R background to obtain the D(-1)-brane effective action deformed by the R-R background. We show that the deformed D(-1)-brane effective action agrees with the instanton effective action in the Omega-background.Comment: 35 pages, no figures, references adde

Average Structures of a Single Knotted Ring Polymer

Two types of average structures of a single knotted ring polymer are studied by Brownian dynamics simulations. For a ring polymer with N segments, its structure is represented by a 3N -dimensional conformation vector consisting of the Cartesian coordinates of the segment positions relative to the center of mass of the ring polymer. The average structure is given by the average conformation vector, which is self-consistently defined as the average of the conformation vectors obtained from a simulation each of which is rotated to minimize its distance from the average conformation vector. From each conformation vector sampled in a simulation, 2N conformation vectors are generated by changing the numbering of the segments. Among the 2N conformation vectors, the one closest to the average conformation vector is used for one type of the average structure. The other type of the averages structure uses all the conformation vectors generated from those sampled in a simulation. In thecase of the former average structure, the knotted part of the average structure is delocalized for small N and becomes localized as N is increased. In the case of the latter average structure, the average structure changes from a double loop structure for small N to a single loop structure for large N, which indicates the localization-delocalization transition of the knotted part.Comment: 15 pages, 19 figures, uses jpsj2.cl

Structure and function of the Rad9-binding region of the DNA-damage checkpoint adaptor TopBP1

TopBP1 is a scaffold protein that coordinates activation of the DNA-damage-checkpoint response by coupling binding of the 9-1-1 checkpoint clamp at sites of ssDNA, to activation of the ATR-ATRIP checkpoint kinase complex. We have now determined the crystal structure of the N-terminal region of human TopBP1, revealing an unexpected triple-BRCT domain structure. The arrangement of the BRCT domains differs significantly from previously described tandem BRCT domain structures, and presents two distinct sites for binding phosphopeptides in the second and third BRCT domains. We show that the site in the second but not third BRCT domain in the N-terminus of TopBP1, provides specific interaction with a phosphorylated motif at pSer387 in Rad9, which can be generated by CK2

N=1/2 Supersymmetric gauge theory in noncommutative space

A formulation of (non-anticommutative) N=1/2 supersymmetric U(N) gauge theory in noncommutative space is studied. We show that at one loop UV/IR mixing occurs. A generalization of Seiberg-Witten map to noncommutative and non-anticommutative superspace is employed to obtain an action in terms of commuting fields at first order in the noncommutativity parameter tetha. This leads to abelian and non-abelian gauge theories whose supersymmetry transformations are local and non-local, respectively.Comment: One reference added, published versio