Cytokinesis in many organisms requires a plasma membrane anchored actomyosin ring, whose contraction facilitates cell division. In yeast and fungi, actomyosin ring constriction is also coordinated ...with division septum assembly. How the actomyosin ring interacts with the plasma membrane and the plasma membrane-localized septum synthesizing machinery remains poorly understood. In Schizosaccharomyces pombe, an attractive model organism to study cytokinesis, the β-1,3-glucan synthase Cps1p / Bgs1p, an integral membrane protein, localizes to the plasma membrane overlying the actomyosin ring and is required for primary septum synthesis. Through a high-dosage suppressor screen we identified an essential gene, sbg1+ (suppressor of beta glucan synthase 1), which suppressed the colony formation defect of Bgs1-defective cps1-191 mutant at higher temperatures. Sbg1p, an integral membrane protein, localizes to the cell ends and to the division site. Sbg1p and Bgs1p physically interact and are dependent on each other to localize to the division site. Loss of Sbg1p results in an unstable actomyosin ring that unravels and slides, leading to an inability to deposit a single contiguous division septum and an important reduction of the β-1,3-glucan proportion in the cell wall, coincident with that observed in the cps1-191 mutant. Sbg1p shows genetic and / or physical interaction with Rga7p, Imp2p, Cdc15p, and Pxl1p, proteins known to be required for actomyosin ring integrity and efficient septum synthesis. This study establishes Sbg1p as a key member of a group of proteins that link the plasma membrane, the actomyosin ring, and the division septum assembly machinery in fission yeast.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The Suzuki‐Miyaura cross‐coupling is one of the most useful synthetic tools to build C−C bonds, but the use of hydroxyaryl halides to direct access hydroxy‐byaryls still remains a challenge. For the ...carbonylative version of the reaction, the synthesis of hydroxybiaryl ketones is commonly solved by protecting group strategies. In this work, we report a protocol of carbonylative Suzuki‐ Miyaura coupling catalysed by a N,P‐PdCl2 complex using aryl halides, various aryl boronic acids and CO. We were able at first to obtain biphenyls, finding a singular reactivity towards 2‐bromophenol, that later was extended to the carbonylation reaction, resulting in methodology that allows the obtention of 2‐hydroxybiaryl ketones, with a functional group tolerance toward amines, alkoxy, ketones, esters, and halides.
It is widely accepted in eukaryotes that the cleavage furrow only initiates after mitosis completion. In fission yeast, cytokinesis requires the synthesis of a septum tightly coupled to cleavage ...furrow ingression. The current cytokinesis model establishes that simultaneous septation and furrow ingression only initiate after spindle breakage and mitosis exit. Thus, this model considers that although Cdk1 is inactivated at early-anaphase, septation onset requires the long elapsed time until mitosis completion and full activation of the Hippo-like SIN pathway. Here, we studied the precise timing of septation onset regarding mitosis by exploiting both the septum-specific detection with the fluorochrome calcofluor and the high-resolution electron microscopy during anaphase and telophase. Contrarily to the existing model, we found that both septum and cleavage furrow start to ingress at early anaphase B, long before spindle breakage, with a slow ingression rate during anaphase B, and greatly increasing after telophase onset. This shows that mitosis and cleavage furrow ingression are not concatenated but simultaneous events in fission yeast. We found that the timing of septation during early anaphase correlates with the cell size and is regulated by the corresponding levels of SIN Etd1 and Rho1. Cdk1 inactivation was directly required for timely septation in early anaphase. Strikingly the reduced SIN activity present after Cdk1 loss was enough to trigger septation by immediately inducing the medial recruitment of the SIN kinase complex Sid2-Mob1. On the other hand, septation onset did not depend on the SIN asymmetry establishment, which is considered a hallmark for SIN activation. These results recalibrate the timing of key cytokinetic events in fission yeast; and unveil a size-dependent control mechanism that synchronizes simultaneous nuclei separation with septum and cleavage furrow ingression to safeguard the proper chromosome segregation during cell division.
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Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
In fungal cells cytokinesis requires coordinated closure of a contractile actomyosin ring (CAR) and synthesis of a special cell wall structure known as the division septum. Many CAR proteins have ...been identified and characterized, but how these molecules interact with the septum synthesis enzymes to form the septum remains unclear. Our genetic study using fission yeast shows that cooperation between the paxillin homolog Pxl1, required for ring integrity, and Bgs1, the enzyme responsible for linear β(1,3)glucan synthesis and primary septum formation, is required for stable anchorage of the CAR to the plasma membrane before septation onset, and for cleavage furrow formation. Thus, lack of Pxl1 in combination with Bgs1 depletion, causes failure of ring contraction and lateral cell wall overgrowth towards the cell lumen without septum formation. We also describe here that Pxl1 concentration at the CAR increases during cytokinesis and that this increase depends on the SH3 domain of the F-BAR protein Cdc15. In consequence, Bgs1 depletion in cells carrying a cdc15ΔSH3 allele causes ring disassembly and septation blockage, as it does in cells lacking Pxl1. On the other hand, the absence of Pxl1 is lethal when Cdc15 function is affected, generating a large sliding of the CAR with deposition of septum wall material along the cell cortex, and suggesting additional functions for both Pxl1 and Cdc15 proteins. In conclusion, our findings indicate that CAR anchorage to the plasma membrane through Cdc15 and Pxl1, and concomitant Bgs1 activity, are necessary for CAR maintenance and septum formation in fission yeast.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
•Extended spectrum β-lactamase producing Escherichia coli were isolated from faeces of 9/53 (17%) healthy dogs in Mexico.•CMY-2 β-lactamase was produced by E. coli isolates from 6/53 (11%) canine ...faecal samples.•Companion animals are potential reservoirs of multiresistant E. coli.•Dissemination of resistance genes by dogs could have important implications in public health.
The presence of extended spectrum β-lactamase (ESBL) and plasmid-mediated AmpC β-lactamase (pAmpC) producing Escherichia coli, along with the mechanisms of antimicrobial resistance and the molecular types of isolates, was investigated in faecal samples from 53 healthy dogs in Mexico. Samples were inoculated on Levine agar plates with 2 µg/mL cefotaxime for recovery of cefotaxime-resistant (CTXR) E. coli. CTXRE. coli isolates were recovered from 9/53 (17%) samples; one isolate was characterised from each positive sample. ESBL producing E. coli isolates were detected in 3/53 (6%) samples; these isolates carried the blaCTX-M-15 gene and one isolate also carried blaSHV-2. These three ESBL-positive E. coli isolates belonged to phylogroup A and sequence types ST617, ST410 or ST3944. The remaining 6/53 (11%) samples contained pAmpC positive isolates; these isolates carried the blaCMY-2 gene, which encodes CMY-2 β-lactamase. These six isolates belonged to phylogroups A (n = 2), B1 (n = 1) and D (n = 3), and sequences types ST1431, ST57, ST93 and ST4565. One CMY-2 β-lactamase positive E. coli isolate of lineage ST93 had the -32 mutation in the chromosomal ampC promoter/attenuator region. Five ESBL/pAmpC positive E. coli isolates carried class 1 integrons (dfrA17-aadA5, aadA and aadA/aadB arrays were detected in three isolates) and one isolate carried a class 2 integron (dfrA12-sat2-aadA1). The aac(6′)Ib-cr, aac(3)-II, qnrB19, tet(A), tet(B), cmlA, and sul3 genes were also detected. All studied isolates showed unrelated PFGE-patterns. To our knowledge, this is the first description of ESBL-producing E. coli and the second of pAmpC-producing E. coli from healthy dogs in America. Our results suggest the potential zoonotic role of dogs in the transmission to humans of ESBL and pAmpC E. coli in the household environment.
P‐pyrrole phosphines (R2Ppyr), in which a pyrrole group is directly bonded to the phosphorus atom, act as monodentate k‐P ligands towards gold(I) center to afford either neutral or cationic ...mononuclear complexes as well as neutral dinuclear complexes. All of these new gold(I) complexes have been structurally characterized and their first uses in catalysis have demonstrated their effectiveness as precatalysts for the enyne cycloisomerization reactions.
P‐pyrrole phosphines (R2Ppyr), in which a pyrrole group is directly bonded to the phosphorus atom, act as monodentate k‐P ligands towards gold(I) center to afford either neutral or cationic mononuclear complexes as well as neutral dinuclear complexes. All of these new gold(I) complexes have been structurally characterized and their first uses in catalysis have demonstrated their effectiveness as precatalysts for the enyne cycloisomerisation reactions.
This study proposes a novel bioinspired metaheuristic simulating how the coronavirus spreads and infects healthy people. From a primary infected individual (patient zero), the coronavirus rapidly ...infects new victims, creating large populations of infected people who will either die or spread infection. Relevant terms such as reinfection probability, super-spreading rate, social distancing measures, or traveling rate are introduced into the model to simulate the coronavirus activity as accurately as possible. The infected population initially grows exponentially over time, but taking into consideration social isolation measures, the mortality rate, and number of recoveries, the infected population gradually decreases. The coronavirus optimization algorithm has two major advantages when compared with other similar strategies. First, the input parameters are already set according to the disease statistics, preventing researchers from initializing them with arbitrary values. Second, the approach has the ability to end after several iterations, without setting this value either. Furthermore, a parallel multivirus version is proposed, where several coronavirus strains evolve over time and explore wider search space areas in less iterations. Finally, the metaheuristic has been combined with deep learning models, to find optimal hyperparameters during the training phase. As application case, the problem of electricity load time series forecasting has been addressed, showing quite remarkable performance.
This work proposes a novel algorithm to forecast big data time series. Based on the well-established Pattern Sequence-based Forecasting algorithm, this new approach has two major contributions to the ...literature. First, the improvement of the original algorithm with respect to the accuracy of predictions, and second, its transformation into the big data context, having reached meaningful results in terms of scalability. The algorithm uses the Apache Spark distributed computation framework and it is a ready-to-use application with few parameters to adjust. Physical and cloud clusters have been used to carry out the experimentation, which consisted in applying the algorithm to real-world data from Uruguay electricity demand.
Time series forecasting has become indispensable for multiple applications and industrial processes. Currently, a large number of algorithms have been developed to forecast time series, all of which ...are suitable depending on the characteristics and patterns to be inferred in each case. In this work, a new algorithm is proposed to predict both univariate and multivariate time series based on a combination of clustering, classification and forecasting techniques. The main goal of the proposed algorithm is first to group windows of time series values with similar patterns by applying a clustering process. Then, a specific forecasting model for each pattern is built and training is only conducted with the time windows corresponding to that pattern. The new algorithm has been designed using a flexible framework that allows the model to be generated using any combination of approaches within multiple machine learning techniques. To evaluate the model, several experiments are carried out using different configurations of the clustering, classification and forecasting methods that the model consists of. The results are analyzed and compared to classical prediction models, such as autoregressive, integrated, moving average and Holt-Winters models, to very recent forecasting methods, including deep, long short-term memory neural networks, and to well-known methods in the literature, such as k nearest neighbors, classification and regression trees, as well as random forest.
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