scholarly journals The In Vitro Characterization of the Role of Rad57 in Rad51 Catalyzed Homologous Recombination Repair

2015 ◽  
Vol 29 (S1) ◽  
Author(s):  
Maria Cusano ◽  
Chioma Ezenduka ◽  
Mark Sweezy
Keyword(s):  
Homologous Recombination ◽  
Homologous Recombination Repair ◽  
In Vitro Characterization ◽  
Recombination Repair

scholarly journals Deficient SUMO Attachment to Flp Recombinase Leads to Homologous Recombination-dependent Hyperamplification of the Yeast 2 μm Circle Plasmid

2009 ◽  
Vol 20 (4) ◽  
pp. 1241-1251 ◽  
Author(s):  
Ling Xiong ◽  
Xiaole L. Chen ◽  
Hannah R. Silver ◽  
Noreen T. Ahmed ◽  
Erica S. Johnson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Homologous Recombination ◽  
Homologous Recombination Repair ◽  
Secondary Effects ◽  
Rolling Circle ◽  
Flp Recombinase ◽  
Recombination Repair ◽  
Sumo Pathway ◽  
Break Induced Replication

Many Saccharomyces cerevisiae mutants defective in the SUMO pathway accumulate elevated levels of the native 2 μm circle plasmid (2 μm). Here we show that accumulation of 2 μm in the SUMO pathway mutants siz1Δ siz2Δ, slx5Δ, and slx8Δ is associated with formation of an aberrant high-molecular-weight (HMW) form of 2 μm. Characterization of this species from siz1Δ siz2Δ showed that it contains tandem copies of the 2 μm sequence as well as single-stranded DNA. Accumulation of this species requires both the 2 μm–encoded Flp recombinase and the cellular homologous recombination repair (HRR) pathway. Importantly, reduced SUMO attachment to Flp is sufficient to induce formation of this species. Our data suggest a model in which Flp that cannot be sumoylated causes DNA damage, whose repair via HRR produces an intermediate that generates tandem copies of the 2 μm sequence. This intermediate may be a rolling circle formed via break-induced replication (BIR), because mutants defective in BIR contain reduced levels of the HMW form. This work also illustrates the importance of using cir° strains when studying mutants that affect the yeast SUMO pathway, to avoid confusing direct functions of the SUMO pathway with secondary effects of 2 μm amplification.

scholarly journals Discovery of Novel Imidazopyridine GSK-3β Inhibitors Supported by Computational Approaches

Molecules ◽  
2020 ◽  
Vol 25 (9) ◽  
pp. 2163
Author(s):  
Rosa Buonfiglio ◽  
Federica Prati ◽  
Martina Bischetti ◽  
Claudia Cavarischia ◽  
Guido Furlotti ◽  
...  
Keyword(s):  
Fine Tuning ◽  
Inhibition Activity ◽  
Design Synthesis ◽  
In Vitro Characterization ◽  
Gsk 3Β ◽  
In Silico Models ◽  
Acidic Hydrogen

The interest of research groups and pharmaceutical companies to discover novel GSK-3β inhibitors has increased over the years considering the involvement of this enzyme in many pathophysiological processes and diseases. Along this line, we recently reported on 1H-indazole-3-carboxamide (INDZ) derivatives 1–6, showing good GSK-3β inhibition activity. However, they suffered from generally poor central nervous system (CNS) permeability. Here, we describe the design, synthesis, and in vitro characterization of novel imidazo[1,5-a]pyridine-1-carboxamide (IMID 1) and imidazo[1,5-a]pyridine-3-carboxamide (IMID 2) compounds (7–18) to overcome such liability. In detail, structure-based approaches and fine-tuning of physicochemical properties guided the design of derivatives 7–18 resulting in ameliorated absorption, distribution, metabolism, and excretion (ADME) properties. A crystal structure of 16 in complex with GSK-3β enzyme (PDB entry 6Y9S) confirmed the in silico models. Despite the nanomolar inhibition activity, the new core compounds showed a reduction in potency with respect to INDZ derivatives 1–6. In this context, Molecular Dynamics (MD) and Quantum Mechanics (QM) based approaches along with NMR investigation helped to rationalize the observed structure activity relationship (SAR). With these findings, the key role of the acidic hydrogen of the central core for a tight interaction within the ATP pocket of the enzyme reflecting in good GSK-3β affinity was demonstrated.

scholarly journals In Vitro Characterization of Peptidoglycan-Associated Lipoprotein (PAL)–Peptidoglycan and PAL–TolB Interactions

1999 ◽  
Vol 181 (20) ◽  
pp. 6306-6311 ◽  
Author(s):  
Emmanuelle Bouveret ◽  
Hélène Bénédetti ◽  
Alain Rigal ◽  
Erwann Loret ◽  
Claude Lazdunski
Keyword(s):  
Outer Membrane ◽  
Cell Envelope ◽  
Membrane Integrity ◽  
Periplasmic Protein ◽  
Multiprotein Complex ◽  
In Vitro Characterization ◽  
In Vitro Interaction

ABSTRACT The Tol-peptidoglycan-associated lipoprotein (PAL) system ofEscherichia coli is a multiprotein complex of the envelope involved in maintaining outer membrane integrity. PAL and the periplasmic protein TolB, two components of this complex, are interacting with each other, and they have also been reported to interact with OmpA and the major lipoprotein, two proteins interacting with the peptidoglycan. All these interactions suggest a role of the Tol-PAL system in anchoring the outer membrane to the peptidoglycan. Therefore, we were interested in better understanding the interaction between PAL and the peptidoglycan. We designed an in vitro interaction assay based on the property of purified peptidoglycan to be pelleted by ultracentrifugation. Using this assay, we showed that a purified PAL protein interacted in vitro with pure peptidoglycan. A peptide competition experiment further demonstrated that the region from residues 89 to 130 of PAL was sufficient to bind the peptidoglycan. Moreover, the fact that this same region of PAL was also binding to TolB suggested that these two interactions were exclusive. Indeed, the TolB-PAL complex appeared not to be associated with the peptidoglycan. This led us to the conclusion that PAL may exist in two forms in the cell envelope, one bound to TolB and the other bound to the peptidoglycan.

Characterization of Autonomic Receptors in Isolated Human Parotid Acinar Cells

1980 ◽  
Vol 59 (2) ◽  
pp. 168-175 ◽  
Author(s):  
John A. Mangos
Keyword(s):  
Adrenergic Receptors ◽  
Acinar Cells ◽  
Cellular Systems ◽  
Parotid Acinar Cells ◽  
In Vitro Characterization ◽  
Muscarinic Cholinergic ◽  
Autonomic Receptors

Isolated human parotid acinar cells have been used for the in vitro characterization of the muscarinic cholinergic and alpha- and beta-adrenergic receptors of these cells. The agonist-antagonist interactions at the receptor level were studied, and the role of the receptor-activated cellular systems in the process of secretion was characterized.

scholarly journals Preparation and In-vitro Characterization of Gastroretentive Floating Tablets of Domperidone

2019 ◽  
Vol 22 (2) ◽  
pp. 170-175 ◽  
Author(s):  
Ikramul Hasan ◽  
Tushar Saha ◽  
Md Selim Reza
Keyword(s):  
Pharmaceutical Journal ◽  
Release Mechanism ◽  
Physical Parameters ◽  
First Order ◽  
In Vitro Characterization ◽  
Time Determination ◽  
Acidic Dissolution

The present investigation was design for domperidone floating table preparation and in-vitro characterization. The ultimate target was increasing gastric retention by means of floatability of the tablet. Hydrophilic cellulosic polymers, Methocel K15M and Methocel K100M were used in this experiment for achieving release controlling property. Sodium bicarbonate played the key role of floatation by generating gas. Direct compression was the method of choice for preparing the tablets. The tablets were evaluated for physical parameters, buoyancy study, total floating time determination and dissolution study. Acidic dissolution medium (0.1N HCl), mimicking the environment of the stomach, was used in USP II apparatus for 12 hours to find out the pattern of drug release. The release mechanism was analyzed by exploring the zero order, first order, Higuchi and Korsmeyer equations. All the physical parameters were within acceptable range and Methocel K100M showed more floating lag time and sustained release property than Methocel K15M. All the formulations showed more than 12 hours floating time. Fourier Transform Infrared Spectroscopy (FTIR) study confirmed the compatibility of the drug with the excipients. Bangladesh Pharmaceutical Journal 22(2): 170-175, 2019

4-Aminobiphenyl inhibits the DNA homologous recombination repair in human liver cells: The role of miR-630 in downregulating RAD18 and MCM8

Toxicology ◽  
2020 ◽  
Vol 440 ◽  
pp. 152441 ◽  
Author(s):  
Heng-Dao Lin ◽  
Fang-Zong Wang ◽  
Chia-Yun Lee ◽  
Chung-Yi Nien ◽  
Yi-Kuan Tseng ◽  
...  
Keyword(s):  
Homologous Recombination ◽  
Human Liver ◽  
Liver Cells ◽  
Homologous Recombination Repair ◽  
Human Liver Cells ◽  
Recombination Repair

Effect of miR-506 on pirarubicin sensitivity and on mesenchymal to epithelial transition and DNA damage homologous recombination repair process in leiomyosarcoma.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. e22503-e22503
Author(s):  
Jilong Yang ◽  
Ting Li ◽  
Feng Li
Keyword(s):  
Dna Damage ◽  
Soft Tissue ◽  
Homologous Recombination ◽  
Drug Sensitivity ◽  
Repair Process ◽  
Homologous Recombination Repair ◽  
Mesenchymal To Epithelial Transition ◽  
Recombination Repair ◽  
Related Proteins

e22503 Background: MicroRNAs has been shown to be involved in many processes of tumorigenesis. Furthermore, miR-506 has been confirmed to be related to the occurrence of epithelial to mesenchymal transition (EMT) and tumor chemoresistance. However, the role of miR-506 in leiomyosarcoma has not yet been reported. Methods: A retrospective review of the 61 Chinese patients diagnosed with soft tissue leiomyosarcoma from January 2000 to December 2012 was investigated. Patient survival trends were examined by Kaplan-Meier analysis and log-rank tests. In vitro experiments were used to determine the biological function and physiological mechanism of drug sensitivity of miR-506. Results: In Chinese leiomyosarcoma patient cohort, we detected a decreased expression of miR-506 in the tumor tissues than that in marginal normal tissues. Tissue microarray analysis detected and confirmed a novel mesenchymal to epithelial transition (MET) in soft tissue leiomyosarcoma, also this MET phenomena was correlated with survival. In vitro, miR-506 inhibited the proliferation, migration, and invasion of leiomyosarcoma cells by promoting the occurrence of MET. Furthermore, RAD51, RAD52, ATM and ATR, which are defined as DNA damage homologous recombination repair related proteins, were highly expressed and associated with worse overall survival of leiomyosarcoma patients. Through targeting MET and DNA damage homologous recombination repair related proteins, miR-506 promoted the sensitivity of leiomyosarcoma cells to Pirarubicin. Conclusions: The current study reveals a novel mechanism that miR-506 increases the drug sensitivity of tumor cells, which are probably in association with the MET and DNA damage homologous recombination repair process.

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