Comparative phosphoproteomic analysis reveals signaling networks regulating monopolar and bipolar cytokinesis.

The successful completion of cytokinesis requires the coordinated activities of diverse cellular components including membranes, cytoskeletal elements and chromosomes that together form partly redundant pathways, depending on the cell type. The biochemical analysis of this process is challenging due to its dynamic and rapid nature. Here, we systematically compared monopolar and bipolar cytokinesis and demonstrated that monopolar cytokinesis is a good surrogate for cytokinesis and it is a well-suited system for global biochemical analysis in mammalian cells. Based on this, we established a phosphoproteomic signature of cytokinesis. More than 10,000 phosphorylation sites were systematically monitored; around 800 of those were up-regulated during cytokinesis. Reconstructing the kinase-substrate interaction network revealed 31 potentially active kinases during cytokinesis. The kinase-substrate network connects proteins between cytoskeleton, membrane and cell cycle machinery. We also found consensus motifs of phosphorylation sites that can serve as biochemical markers specific to cytokinesis. Beyond the kinase-substrate network, our reconstructed signaling network suggests that combination of sumoylation and phosphorylation may regulate monopolar cytokinesis specific signaling pathways. Our analysis provides a systematic approach to the comparison of different cytokinesis types to reveal alternative ways and a global overview, in which conserved genes work together and organize chromatin and cytoplasm during cytokinesis.EMBO (European Molecular Biology Organization) Installation Grant; Young Scientist Award Program BAGEP of the Science Academy (Turkey); TUBITAK-Marie Curie Co-funded Brain Circulation Schem

Accurate MS-based Rab10 phosphorylation stoichiometry determination as readout for LRRK2 activity in Parkinson's disease

Pathogenic mutations in the Leucine-rich repeat kinase 2 (LRRK2) increase its activity leading to increased phosphorylation of Rab proteins. Here we introduce a sensitive and accurate assay to measure increased phospho Rab levels using synthetic stable isotope-labeled analogues for both phosphorylated and non-phosphorylated tryptic peptides surrounding Rab10-Thr73. Compared to healthy controls, carriers of mutated LRRK2 had 1.9-fold and those with VPS35 mutation 3.7-fold increased pRab10 levels in their neutrophils. Our generic MS-based assay helps to stratify PD patient and determine LRRK2 inhibitor efficiency in clinical trials. Pathogenic mutations in the Leucine-rich repeat kinase 2 (LRRK2) are the predominant genetic cause of Parkinson's disease (PD). They increase its activity, resulting in augmented Rab10-Thr73 phosphorylation and conversely, LRRK2 inhibition decreases pRab10 levels. Currently, there is no assay to quantify pRab10 levels for drug target engagement or patient stratification. To meet this challenge, we developed an high accuracy and sensitivity targeted mass spectrometry (MS)-based assay for determining Rab10-Thr73 phosphorylation stoichiometry in human samples. It uses synthetic stable isotope-labeled (SIL) analogues for both phosphorylated and nonphosphorylated tryptic peptides surrounding Rab10-Thr73 to directly derive the percentage of Rab10 phosphorylation from attomole amounts of the endogenous phosphopeptide. The SIL and the endogenous phosphopeptides are separately admitted into an Orbitrap analyzer with the appropriate injection times. We test the reproducibility of our assay by determining Rab10-Thr73 phosphorylation stoichiometry in neutrophils of LRRK2 mutation carriers before and after LRRK2 inhibition. Compared with healthy controls, the PD predisposing mutation carriers LRRK2 G2019S and VPS35 D620N display 1.9-fold and 3.7-fold increased pRab10 levels, respectively. Our generic MS-based assay further establishes the relevance of pRab10 as a prognostic PD marker and is a powerful tool for determining LRRK2 inhibitor efficacy and for stratifying PD patients for LRRK2 inhibitor treatment.Acknowledgments: We thank Sabine Suppmann, Leopold Urich, Stephan Uebel, Stefan Pettera, Martin Spitaler, Nagarjuna Nagaraj, Victoria Sanchez and Antonio Piras from the MPIB Biochemistry Core Facility

DNase Sda1 Allows Invasive M1T1 Group A Streptococcus to Prevent TLR9-Dependent Recognition

Group A Streptococcus (GAS) has developed a broad arsenal of virulence factors that serve to circumvent host defense mechanisms. The virulence factor DNase Sda1 of the hyperinvasive M1T1 GAS clone degrades DNA-based neutrophil extracellular traps allowing GAS to escape extracellular killing. TLR9 is activated by unmethylated CpG-rich bacterial DNA and enhances innate immune resistance. We hypothesized that Sda1 degradation of bacterial DNA could alter TLR9-mediated recognition of GAS by host innate immune cells. We tested this hypothesis using a dual approach: loss and gain of function of DNase in isogenic GAS strains and presence and absence of TLR9 in the host. Either DNA degradation by Sda1 or host deficiency of TLR9 prevented GAS induced IFN-α and TNF-α secretion from murine macrophages and contributed to bacterial survival. Similarly, in a murine necrotizing fasciitis model, IFN-α and TNF-α levels were significantly decreased in wild type mice infected with GAS expressing Sda1, whereas no such Sda1-dependent effect was seen in a TLR9-deficient background. Thus GAS Sda1 suppressed both the TLR9-mediated innate immune response and macrophage bactericidal activity. Our results demonstrate a novel mechanism of bacterial innate immune evasion based on autodegradation of CpG-rich DNA by a bacterial DNase

Cassini Radio Science

Cassini radio science investigations will be conducted both during the cruise (gravitational wave and conjunction experiments) and the Saturnian tour of the mission (atmospheric and ionospheric occultations, ring occultations, determinations of masses and gravity fields). New technologies in the construction of the instrument, which consists of a portion on-board the spacecraft and another portion on the ground, including the use of the Ka-band signal in addition to that of the S- and X-bands, open opportunities for important discoveries in each of the above scientific areas, due to increased accuracy, resolution, sensitivity, and dynamic range.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/43765/1/11214_2004_Article_1436.pd

Usage of geographic information systems based modelling in management and planning of disaster planning

Информационные системы и технологии в экологии и здравоохраненииNatural disasters are a part of life and may have extremely destructive effects if necessary precautions are not taken. Geographic Information Systems (GIS) are a part of the management systems that present both verbal and graphic data, spatial analyses and other bases to those in positions of decision-making to support in their decisions and planning. GIS enables the storage and questioning of data concerning the current situation and at the same time ensures making analyses, inquiries and predictions for the future. In this study, the natural disasters that took place in our country between the years 1970 and 2015 were dealt with. Data belonging to DEMA (Disaster and Emergency Management Authority) were compiled and they were transferred to the GIS database. Using these data, maps of geographical regions which natural disasters affected according to the frequency of occurrence were created in GIS environment. What kind of disasters took place in what geographical regions were revealed through thematic maps obtained as a consequence of spatial analyses = Стихийные бедствия являются частью жизни и могут иметь чрезвычайно разрушительные последствия, если не будут приняты необходимые меры предосторожности. Географические информационные системы (ГИС) являются частью систем управления, которые представляют как устные, так и графические данные, пространственные анализы и другие базы тем, кто находится на руководящих должностях, для поддержки их решений и планирования. ГИС позволяет хранить и подвергать сомнению данные о текущей ситуации и в то же время обеспечивает проведение анализов, запросов и прогнозов на будущее. В этом исследовании были рассмотрены стихийные бедствия, имевшие место в нашей стране в период между 1970 и 2015 г. Данные, принадлежащие DEMA (Управление по чрезвычайным ситуациям и чрезвычайным ситуациям), были скомпилированы, и они были переданы в базу данных ГИС. Используя эти данные, в географической среде были созданы карты географических регионов, в которых стихийные бедствия пострадали в зависимости от частоты их возникновения. Какие бедствия имели место в каких географических регионах были выявлены с помощью тематических карт, полученных в результате пространственного анализ

Usage of geographic information systems based modelling in management and planning of disaster planning

Информационные системы и технологии в экологии и здравоохраненииNatural disasters are a part of life and may have extremely destructive effects if necessary precautions are not taken. Geographic Information Systems (GIS) are a part of the management systems that present both verbal and graphic data, spatial analyses and other bases to those in positions of decision-making to support in their decisions and planning. GIS enables the storage and questioning of data concerning the current situation and at the same time ensures making analyses, inquiries and predictions for the future. In this study, the natural disasters that took place in our country between the years 1970 and 2015 were dealt with. Data belonging to DEMA (Disaster and Emergency Management Authority) were compiled and they were transferred to the GIS database. Using these data, maps of geographical regions which natural disasters affected according to the frequency of occurrence were created in GIS environment. What kind of disasters took place in what geographical regions were revealed through thematic maps obtained as a consequence of spatial analyses = Стихийные бедствия являются частью жизни и могут иметь чрезвычайно разрушительные последствия, если не будут приняты необходимые меры предосторожности. Географические информационные системы (ГИС) являются частью систем управления, которые представляют как устные, так и графические данные, пространственные анализы и другие базы тем, кто находится на руководящих должностях, для поддержки их решений и планирования. ГИС позволяет хранить и подвергать сомнению данные о текущей ситуации и в то же время обеспечивает проведение анализов, запросов и прогнозов на будущее. В этом исследовании были рассмотрены стихийные бедствия, имевшие место в нашей стране в период между 1970 и 2015 г. Данные, принадлежащие DEMA (Управление по чрезвычайным ситуациям и чрезвычайным ситуациям), были скомпилированы, и они были переданы в базу данных ГИС. Используя эти данные, в географической среде были созданы карты географических регионов, в которых стихийные бедствия пострадали в зависимости от частоты их возникновения. Какие бедствия имели место в каких географических регионах были выявлены с помощью тематических карт, полученных в результате пространственного анализ