scholarly journals Redox controls RecA protein activity via reversible oxidation of its methionine residues

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Camille Henry ◽  
Laurent Loiseau ◽  
Alexandra Vergnes ◽  
Didier Vertommen ◽  
Angela Mérida-Floriano ◽  
...  
Keyword(s):  
Dna Recombination ◽  
Reca Protein ◽  
Methionine Sulfoxide ◽  
Mass Spectrometry Analysis ◽  
Loss Of Function ◽  
Protein Activity ◽  
Spectrometry Analysis ◽  
Methionine Sulfoxide Reductases ◽  
Methionine Residues ◽  
Biochemical Analyses

Reactive oxygen species (ROS) cause damage to DNA and proteins. Here we report that the RecA recombinase is itself oxidized by ROS. Genetic and biochemical analyses revealed that oxidation of RecA altered its DNA repair and DNA recombination activities. Mass spectrometry analysis showed that exposure to ROS converted 4 out of 9 Met residues of RecA to methionine sulfoxide. Mimicking oxidation of Met35 by changing it for Gln caused complete loss of function whereas mimicking oxidation of Met164 resulted in constitutive SOS activation and loss of recombination activity. Yet, all ROS-induced alterations of RecA activity were suppressed by methionine sulfoxide reductases MsrA and MsrB. These findings indicate that under oxidative stress, MsrA/B is needed for RecA homeostasis control. The implication is that, besides damaging DNA structure directly, ROS prevent repair of DNA damage by hampering RecA activity.

scholarly journals In Vivo Effects of Methionine Sulfoxide Reductase Deficiency in Drosophila melanogaster

Antioxidants ◽  
2018 ◽  
Vol 7 (11) ◽  
pp. 155 ◽  
Author(s):  
Lindsay Bruce ◽  
Diana Singkornrat ◽  
Kelsey Wilson ◽  
William Hausman ◽  
Kelli Robbins ◽  
...  
Keyword(s):  
Methionine Sulfoxide ◽  
Methionine Sulfoxide Reductase ◽  
Model Organisms ◽  
Methionine Sulfoxide Reductase A ◽  
Methionine Sulfoxide Reductases ◽  
Methionine Residues ◽  
Oxidation Of Methionine ◽  
And Function ◽  
In Vivo Effects

The deleterious alteration of protein structure and function due to the oxidation of methionine residues has been studied extensively in age-associated neurodegenerative disorders such as Alzheimer’s and Parkinson’s Disease. Methionine sulfoxide reductases (MSR) have three well-characterized biological functions. The most commonly studied function is the reduction of oxidized methionine residues back into functional methionine thus, often restoring biological function to proteins. Previous studies have successfully overexpressed and silenced MSR activity in numerous model organisms correlating its activity to longevity and oxidative stress. In the present study, we have characterized in vivo effects of MSR deficiency in Drosophila. Interestingly, we found no significant phenotype in animals lacking either methionine sulfoxide reductase A (MSRA) or methionine sulfoxide reductase B (MSRB). However, Drosophila lacking any known MSR activity exhibited a prolonged larval third instar development and a shortened lifespan. These data suggest an essential role of MSR in key biological processes.

scholarly journals Preservation of collagen in the soft tissues of frozen mammoths

2021 ◽  
Author(s):  
Shunji Hattori ◽  
Tomomi Kiriyama-Tanaka ◽  
Masashi Kusubata ◽  
Yuki Taga ◽  
Testuya Ebihara ◽  
...  
Keyword(s):  
Soft Tissues ◽  
Mass Spectrometry Analysis ◽  
Fibrillar Structure ◽  
Collagen Molecule ◽  
Type I ◽  
Spectrometry Analysis ◽  
Type Iv ◽  
Cross Striation ◽  
Main Protein ◽  
Biochemical Analyses

We investigated the characteristics of extracellular matrix (ECM) in the soft tissue of two frozen baby woolly mammoths ( Mammuthus primigenius ) that died and were buried in Siberian permafrost approximately 40,000 years ago. Morphological and biochemical analyses of mammoth lung and liver demonstrated that those soft tissues were preserved at the gross anatomical and histological levels. The ultrastructure of ECM components, namely a fibrillar structure with a collagen-characteristic pattern of cross-striation, was clearly visible with transmission and scanning electron microscopy. Type I and type IV collagens were detected by immunohistochemical observation. Quantitative amino acid analysis of liver and lung tissues of the baby mammoths indicated that collagenous protein is selectively preserved in these tissues as a main protein. Type I and type III collagens were detected as major components by means of liquid chromatography–mass spectrometry analysis after digestion with trypsin. These results indicate that the triple helical collagen molecule, which is resistant to proteinase digestion, has been preserved in the soft tissues of these frozen mammoths for 40,000 years.

scholarly journals RACK1 associates with RNA-binding proteins Vigilin and SERBP1 to control dengue virus replication

2021 ◽  
Author(s):  
Alexis Brugier ◽  
Mohamed-Lamine Hafirassou ◽  
Marie Pourcelot ◽  
Morgane Baldaccini ◽  
Laurine Couture ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Binding Proteins ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Ribosomal Subunit ◽  
Denv Infection ◽  
Mass Spectrometry Analysis ◽  
Macromolecular Complex ◽  
Loss Of Function ◽  
Spectrometry Analysis

Dengue virus (DENV), a re-emerging virus transmitted by Aedes mosquitoes, causes severe pathogenesis in humans. No effective treatment is available against this virus. We recently identified the scaffold protein RACK1 as a component of the DENV replication complex, a macromolecular complex essential for viral genome amplification. Here, we show that RACK1 is important for DENV infection. RACK1 mediates DENV replication through binding to the 40S ribosomal subunit. Mass spectrometry analysis of RACK1 partners coupled to a loss-of-function screen identified the RNA binding proteins Vigilin and SERBP1 as DENV host dependency factors. Vigilin and SERBP1 interact with DENV viral RNA (vRNA), forming a ternary complex with RACK1 to mediate viral replication. Overall, our results indicate that RACK1 recruits Vigilin and SERBP1, linking the DENV vRNA to the translation machinery for optimal translation and replication.

scholarly journals GAS CHROMATOGRAPHY-MASS SPECTROMETRY ANALYSIS OF VIBURNUM OPULUS (L) EXTRACT AND ITS TOXICITY STUDIES IN RATS

2017 ◽  
Vol 10 (6) ◽  
pp. 383 ◽  
Author(s):  
Abiodun Humphrey Adebayo ◽  
Aristotle Balade ◽  
Omolara Faith Yakubu
Keyword(s):  
Gas Chromatography ◽  
Leaf Extract ◽  
Density Lipoprotein ◽  
Mass Spectrometry Analysis ◽  
Gas Chromatography Mass Spectrometry ◽  
Water Control ◽  
Viburnum Opulus ◽  
Spectrometry Analysis ◽  
Water Fractions ◽  
Biochemical Analyses

Objectives: This study was aimed at establishing the antimicrobial and phytochemical profiles of Viburnum opulus (L) as well as the safety potential of the extract in albino Wistar rats.Methods: Ethanol, n-hexane, ethyl acetate, butanol and water fractions were prepared for both phytochemical assessment using gas chromatography-mass spectrum analysis (GC-MS). Five groups of seven rats were used for the study. Group A received distilled water (control), while groups B to E were treated respectively with 250, 500, 1000 and 1500 mg/kg body weight of V. opulus extract by abdominal canulisation for 28 days. Blood samples were obtained for biochemical analyses and the liver tissues were further processed for histological studies.Results: The GC-MS spectra revealed the existence of various phytoconstituents such as neophytadiene, germaciene, caryophyllene among others. High density lipoprotein and albumin were significantly (p<0.05) elevated in animals administered with 500, 1000 and 1500 mg/kg bw of the leaf extract. Ethanol, butanol and water fractions of the leaf of V. opulus showed antimicrobial action against most of the organisms used in this study.Conclusion: The result indicates the V. opulus leaf extract contains a wild range of fatty acids and heterocyclic compounds with antimicrobial efficacy and no hepatic damage.

scholarly journals Methionine Redox Homeostasis in Protein Quality Control

2021 ◽  
Vol 8 ◽  
Author(s):  
Laurent Aussel ◽  
Benjamin Ezraty
Keyword(s):  
Oxidative Stress ◽  
Quality Control ◽  
Protein Function ◽  
Protein Quality ◽  
Redox Homeostasis ◽  
Protein Quality Control ◽  
Methionine Sulfoxide ◽  
Methionine Sulfoxide Reductases ◽  
Methionine Residues ◽  
And Function

Bacteria live in different environments and are subject to a wide variety of fluctuating conditions. During evolution, they acquired sophisticated systems dedicated to maintaining protein structure and function, especially during oxidative stress. Under such conditions, methionine residues are converted into methionine sulfoxide (Met-O) which can alter protein function. In this review, we focus on the role in protein quality control of methionine sulfoxide reductases (Msr) which repair oxidatively protein-bound Met-O. We discuss our current understanding of the importance of Msr systems in rescuing protein function under oxidative stress and their ability to work in coordination with chaperone networks. Moreover, we highlight that bacterial chaperones, like GroEL or SurA, are also targeted by oxidative stress and under the surveillance of Msr. Therefore, integration of methionine redox homeostasis in protein quality control during oxidative stress gives a complete picture of this bacterial adaptive mechanism.

scholarly journals Preservation of collagen in the soft tissues of frozen mammoths

PLoS ONE ◽  
2021 ◽  
Vol 16 (10) ◽  
pp. e0258699
Author(s):  
Shunji Hattori ◽  
Tomomi Kiriyama-Tanaka ◽  
Masashi Kusubata ◽  
Yuki Taga ◽  
Testuya Ebihara ◽  
...  
Keyword(s):  
Soft Tissues ◽  
Mass Spectrometry Analysis ◽  
Fibrillar Structure ◽  
Collagen Molecule ◽  
Type I ◽  
Spectrometry Analysis ◽  
Type Iv ◽  
Cross Striation ◽  
Main Protein ◽  
Biochemical Analyses

We investigated the characteristics of extracellular matrix (ECM) in the soft tissue of two frozen baby woolly mammoths (Mammuthus primigenius) that died and were buried in Siberian permafrost approximately 40,000 years ago. Morphological and biochemical analyses of mammoth lung and liver demonstrated that those soft tissues were preserved at the gross anatomical and histological levels. The ultrastructure of ECM components, namely a fibrillar structure with a collagen-characteristic pattern of cross-striation, was clearly visible with transmission and scanning electron microscopy. Type I and type IV collagens were detected by immunohistochemical observation. Quantitative amino acid analysis of liver and lung tissues of the baby mammoths indicated that collagenous protein is selectively preserved in these tissues as a main protein. Type I and type III collagens were detected as major components by means of liquid chromatography–mass spectrometry analysis after digestion with trypsin. These results indicate that the triple helical collagen molecule, which is resistant to proteinase digestion, has been preserved in the soft tissues of these frozen mammoths for 40,000 years.

scholarly journals Loss of Function of the Gene Encoding the Histone Methyltransferase KMT2D Leads to Deregulation of Mitochondrial Respiration

Cells ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 1685
Author(s):  
Consiglia Pacelli ◽  
Iolanda Adipietro ◽  
Natascia Malerba ◽  
Gabriella Maria Squeo ◽  
Claudia Piccoli ◽  
...  
Keyword(s):  
Mitochondrial Respiratory Chain ◽  
Significant Inhibition ◽  
Metabolic Reprogramming ◽  
Mass Spectrometry Analysis ◽  
Metabolic Phenotype ◽  
Glycolytic Flux ◽  
Epigenetic Mark ◽  
Loss Of Function ◽  
Respiratory Chain Complexes ◽  
Spectrometry Analysis

KMT2D encodes a methyltransferase responsible for histone 3 lysine 4 (H3K4) mono-/di-methylation, an epigenetic mark correlated with active transcription. Here, we tested the hypothesis that KMT2D pathogenic loss-of-function variants, which causes the Kabuki syndrome type 1, could affect the mitochondrial metabolic profile. By using Seahorse technology, we showed a significant reduction of the mitochondrial oxygen consumption rate as well as a reduction of the glycolytic flux in both Kmt2d knockout MEFs and skin fibroblasts of Kabuki patients harboring heterozygous KMT2D pathogenic variants. Mass-spectrometry analysis of intermediate metabolites confirmed alterations in the glycolytic and TCA cycle pathways. The observed metabolic phenotype was accompanied by a significant increase in the production of reactive oxygen species. Measurements of the specific activities of the mitochondrial respiratory chain complexes revealed significant inhibition of CI (NADH dehydrogenase) and CIV (cytochrome c oxidase); this result was further supported by a decrease in the protein content of both complexes. Finally, we unveiled an impaired oxidation of glucose and larger reliance on long-chain fatty acids oxidation. Altogether, our findings clearly indicate a rewiring of the mitochondrial metabolic phenotype in the KMT2D-null or loss-of-function context that might contribute to the development of Kabuki disease, and represents metabolic reprogramming as a potential new therapeutic approach.

scholarly journals Reduced carboxylesterase 1 is associated with endothelial injury in methamphetamine-induced pulmonary arterial hypertension

2017 ◽  
Vol 313 (2) ◽  
pp. L252-L266 ◽  
Author(s):  
Mark E. Orcholski ◽  
Artyom Khurshudyan ◽  
Elya A. Shamskhou ◽  
Ke Yuan ◽  
Ian Y. Chen ◽  
...  
Keyword(s):  
Pulmonary Arterial Hypertension ◽  
Arterial Hypertension ◽  
Mass Spectrometry Analysis ◽  
Loss Of Function ◽  
Sequencing Data ◽  
Spectrometry Analysis ◽  
Whole Exome Sequencing Data ◽  
Pulmonary Arterial ◽  
Carboxylesterase 1 ◽  
Induced Apoptosis

Pulmonary arterial hypertension is a complication of methamphetamine use (METH-PAH), but the pathogenic mechanisms are unknown. Given that cytochrome P450 2D6 (CYP2D6) and carboxylesterase 1 (CES1) are involved in metabolism of METH and other amphetamine-like compounds, we postulated that loss of function variants could contribute to METH-PAH. Although no difference in CYP2D6 expression was seen by lung immunofluorescence, CES1 expression was significantly reduced in endothelium of METH-PAH microvessels. Mass spectrometry analysis showed that healthy pulmonary microvascular endothelial cells (PMVECs) have the capacity to both internalize and metabolize METH. Furthermore, whole exome sequencing data from 18 METH-PAH patients revealed that 94.4% of METH-PAH patients were heterozygous carriers of a single nucleotide variant (SNV; rs115629050) predicted to reduce CES1 activity. PMVECs transfected with this CES1 variant demonstrated significantly higher rates of METH-induced apoptosis. METH exposure results in increased formation of reactive oxygen species (ROS) and a compensatory autophagy response. Compared with healthy cells, CES1-deficient PMVECs lack a robust autophagy response despite higher ROS, which correlates with increased apoptosis. We propose that reduced CES1 expression/activity could promote development of METH-PAH by increasing PMVEC apoptosis and small vessel loss.

The role of mass spectrometry analysis in bacterial effector characterization

2017 ◽  
Vol 474 (16) ◽  
pp. 2779-2784 ◽  
Author(s):  
Nichollas E. Scott ◽  
Elizabeth L. Hartland
Keyword(s):  
Mass Spectrometry ◽  
Cell Biology ◽  
Critical Role ◽  
Effector Proteins ◽  
Mass Spectrometry Analysis ◽  
Protein Activity ◽  
Post Translational Modifications ◽  
Spectrometry Analysis ◽  
New Generation

Many secreted bacterial effector proteins play a critical role in host–pathogen interactions by mediating a variety of post-translational modifications, some of which do not occur natively within the eukaryotic proteome. The characterization of bacterial effector protein activity remains an important step to understanding the subversion of host cell biology during pathogen infection and although molecular biology and immunochemistry remain critical tools for gaining insights into bacterial effector functions, increasingly mass spectrometry (MS) and proteomic approaches are also playing an indispensable role. The focus of this editorial is to highlight the strengths of specific MS approaches and their utility for the characterization of bacterial effector activity. With the capability of new generation MS instrumentation, MS-based technologies can provide information that is inaccessible using traditional molecular or immunochemical approaches.

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