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Elucidating the Role of the Escherichia Coli RarA Protein at the Interface of DNA Replication and DNA Repair

Elucidating the Role of the Escherichia Coli RarA Protein at the Interface of DNA Replication and DNA Repair PDF Author: Tyler H. Stanage
Publisher:
ISBN:
Category :
Languages : en
Pages : 0

Book Description
In each cell cycle, the replisome encounters replication blocks that include DNA lesions, tightly bound DNA-protein complexes, and breaks in the DNA. In Escherichia coli, many pathways have evolved to recognize, respond to, and repair these replication blocks so that replication can be completed in a timely and accurate manner. The highly conserved RarA (Replication associated recombination protein A) protein family has been implicated in the faithful resolution of these replication conflicts. Despite high conservation and extensive work conducted on this protein family, the overall function of RarA remains enigmatic. Here, we carefully describe two novel DNA-dependent functions for the E. coli RarA protein. First, we demonstrated that RarA binds to and hydrolyzes ATP at double-stranded DNA ends in order to generate single-stranded DNA flaps. Second, using a single-molecule reconstituted DNA replication assay, we show that RarA generated single-stranded DNA gaps behind active replisomes in an ATP-dependent manner. This activity is achieved by RarA disengaging DNA polymerase III cores from their [beta]2 processivity clamps. In vivo, we demonstrate that RarA--presumably using its gap creation activity--generates suitable substrates for post-replication repair pathways including RecFOR-mediated homologous recombination and translesion DNA synthesis. Loss of this function results in impaired growth and varied DNA replication phenotypes. Overexpression of rarA is toxic due to its ATPase activity and the resulting overuse of mutagenic TLS polymerases. Together, these results further elucidate the role of the RarA protein family at the interface of DNA replication and DNA repair.

Elucidating the Role of the Escherichia Coli RarA Protein at the Interface of DNA Replication and DNA Repair

Elucidating the Role of the Escherichia Coli RarA Protein at the Interface of DNA Replication and DNA Repair PDF Author: Tyler H. Stanage
Publisher:
ISBN:
Category :
Languages : en
Pages : 0

Book Description
In each cell cycle, the replisome encounters replication blocks that include DNA lesions, tightly bound DNA-protein complexes, and breaks in the DNA. In Escherichia coli, many pathways have evolved to recognize, respond to, and repair these replication blocks so that replication can be completed in a timely and accurate manner. The highly conserved RarA (Replication associated recombination protein A) protein family has been implicated in the faithful resolution of these replication conflicts. Despite high conservation and extensive work conducted on this protein family, the overall function of RarA remains enigmatic. Here, we carefully describe two novel DNA-dependent functions for the E. coli RarA protein. First, we demonstrated that RarA binds to and hydrolyzes ATP at double-stranded DNA ends in order to generate single-stranded DNA flaps. Second, using a single-molecule reconstituted DNA replication assay, we show that RarA generated single-stranded DNA gaps behind active replisomes in an ATP-dependent manner. This activity is achieved by RarA disengaging DNA polymerase III cores from their [beta]2 processivity clamps. In vivo, we demonstrate that RarA--presumably using its gap creation activity--generates suitable substrates for post-replication repair pathways including RecFOR-mediated homologous recombination and translesion DNA synthesis. Loss of this function results in impaired growth and varied DNA replication phenotypes. Overexpression of rarA is toxic due to its ATPase activity and the resulting overuse of mutagenic TLS polymerases. Together, these results further elucidate the role of the RarA protein family at the interface of DNA replication and DNA repair.

Elucidating the Role of E. Coli Rara Protein in Dna Replication, Recombination, and Repair Processes

Elucidating the Role of E. Coli Rara Protein in Dna Replication, Recombination, and Repair Processes PDF Author: Kanika Jain (Ph.D.)
Publisher:
ISBN:
Category :
Languages : en
Pages : 347

Book Description
DNA replication, indispensable to the survival and reproduction of all living organisms, is a highly coordinated and complex process. The progress of replication is regularly challenged by extrinsic and intrinsic barriers. These include DNA lesions produced by reactive oxygen species (ROS) or other DNA damaging agents and protein-DNA complexes. Encounters with such barriers can result in replication fork stalling or collapse. Although estimates vary, replication forks in bacteria may stall or collapse as often as once per cell generation during normal growth conditions. Failure to rescue this stalled replication fork may result in genome instability, tumorigenesis, and cell death. However, numerous pathways exist for repairing and reactivating replication forks and these pathways are crucial for maintaining genome stability and cell viability. Bacterial "Replication associated recombination protein A" (RarA) and related eukaryotic enzymes are implicated in the maintenance of genome instability. The RarA protein family (RarA, Mgs1, and WRNP1) is among the most widespread and conserved of any family with a putative role in DNA repair. Despite high conservation and extensive work conducted on this protein family, the overall function of RarA remains enigmatic. We show that the RarA protein plays a prominent role in intermolecular recombination involving relatively short (

DNA Replication, Recombination, and Repair

DNA Replication, Recombination, and Repair PDF Author: Fumio Hanaoka
Publisher: Springer
ISBN: 443155873X
Category : Science
Languages : en
Pages : 548

Book Description
This book is a comprehensive review of the detailed molecular mechanisms of and functional crosstalk among the replication, recombination, and repair of DNA (collectively called the "3Rs") and the related processes, with special consciousness of their biological and clinical consequences. The 3Rs are fundamental molecular mechanisms for organisms to maintain and sometimes intentionally alter genetic information. DNA replication, recombination, and repair, individually, have been important subjects of molecular biology since its emergence, but we have recently become aware that the 3Rs are actually much more intimately related to one another than we used to realize. Furthermore, the 3R research fields have been growing even more interdisciplinary, with better understanding of molecular mechanisms underlying other important processes, such as chromosome structures and functions, cell cycle and checkpoints, transcriptional and epigenetic regulation, and so on. This book comprises 7 parts and 21 chapters: Part 1 (Chapters 1–3), DNA Replication; Part 2 (Chapters 4–6), DNA Recombination; Part 3 (Chapters 7–9), DNA Repair; Part 4 (Chapters 10–13), Genome Instability and Mutagenesis; Part 5 (Chapters 14–15), Chromosome Dynamics and Functions; Part 6 (Chapters 16–18), Cell Cycle and Checkpoints; Part 7 (Chapters 19–21), Interplay with Transcription and Epigenetic Regulation. This volume should attract the great interest of graduate students, postdoctoral fellows, and senior scientists in broad research fields of basic molecular biology, not only the core 3Rs, but also the various related fields (chromosome, cell cycle, transcription, epigenetics, and similar areas). Additionally, researchers in neurological sciences, developmental biology, immunology, evolutionary biology, and many other fields will find this book valuable.

Molecular Diagnostics: Promises and Possibilities

Molecular Diagnostics: Promises and Possibilities PDF Author: Mousumi Debnath
Publisher: Springer Science & Business Media
ISBN: 9048132614
Category : Medical
Languages : en
Pages : 527

Book Description
A rapid development in diverse areas of molecular biology and genetic engineering resulted in emergence of variety of tools. These tools are not only applicable to basic researches being carried out world over, but also exploited for precise detection of abnormal conditions in plants, animals and human body. Although a basic researcher is well versed with few techniques used by him/her in the laboratory, they may not be well acquainted with methodologies, which can be used to work out some of their own research problems. The picture is more blurred when the molecular diagnostic tools are to be used by physicians, scientists and technicians working in diagnostic laboratories in hospitals, industry and academic institutions. Since many of them are not trained in basics of these methods, they come across several gray areas in understanding of these tools. The accurate application of molecular diagnostic tools demands in depth understanding of the methodology for precise detection of the abnormal condition of living body. To meet the requirements of a good book on molecular diagnostics of students, physicians, scientists working in agricultural, veterinary, medical and pharmaceutical sciences, it needs to expose the reader lucidly to: Give basic science behind commonly used tools in diagnostics Expose the readers to detailed applications of these tools and Make them aware the availability of such diagnostic tools The book will attract additional audience of pathologists, medical microbiologists, pharmaceutical sciences, agricultural scientists and veterinary doctors if the following topics are incorporated at appropriate places in Unit II or separately as a part of Unit-III in the book. Molecular diagnosis of diseases in agricultural crops Molecular diagnosis of veterinary diseases. Molecular epidemiology, which helps to differentiate various epidemic strains and sources of disease outbreaks. Even in different units of the same hospital, the infections could be by different strains of the same species and the information becomes valuable for infection control strategies. Drug resistance is a growing problem for bacterial, fungal and parasitic microbes and the molecular biology tools can help to detect the drug resistance genes without the cultivation and in vitro sensitivity testing. Molecular diagnostics offers faster help in the selection of the proper antibiotic for the treatment of tuberculosis, which is a major problem of the in the developing world. The conventional culture and drug sensitivity testing of tuberculosis bacilli is laborious and time consuming, whereas molecular diagnosis offers rapid drug resistant gene detection even from direct clinical samples. The same approach for HIV, malaria and many more diseases needs to be considered. Molecular diagnostics in the detection of diseases during foetal life is an upcoming area in the foetal medicine in case of genetic abnormalities and infectious like TORCH complex etc. The book will be equally useful to students, scientists and professionals working in the field of molecular diagnostics.

The WEIRDest People in the World

The WEIRDest People in the World PDF Author: Joseph Henrich
Publisher: Farrar, Straus and Giroux
ISBN: 0374710457
Category : Psychology
Languages : en
Pages : 420

Book Description
A New York Times Notable Book of 2020 A Bloomberg Best Non-Fiction Book of 2020 A Behavioral Scientist Notable Book of 2020 A Human Behavior & Evolution Society Must-Read Popular Evolution Book of 2020 A bold, epic account of how the co-evolution of psychology and culture created the peculiar Western mind that has profoundly shaped the modern world. Perhaps you are WEIRD: raised in a society that is Western, Educated, Industrialized, Rich, and Democratic. If so, you’re rather psychologically peculiar. Unlike much of the world today, and most people who have ever lived, WEIRD people are highly individualistic, self-obsessed, control-oriented, nonconformist, and analytical. They focus on themselves—their attributes, accomplishments, and aspirations—over their relationships and social roles. How did WEIRD populations become so psychologically distinct? What role did these psychological differences play in the industrial revolution and the global expansion of Europe during the last few centuries? In The WEIRDest People in the World, Joseph Henrich draws on cutting-edge research in anthropology, psychology, economics, and evolutionary biology to explore these questions and more. He illuminates the origins and evolution of family structures, marriage, and religion, and the profound impact these cultural transformations had on human psychology. Mapping these shifts through ancient history and late antiquity, Henrich reveals that the most fundamental institutions of kinship and marriage changed dramatically under pressure from the Roman Catholic Church. It was these changes that gave rise to the WEIRD psychology that would coevolve with impersonal markets, occupational specialization, and free competition—laying the foundation for the modern world. Provocative and engaging in both its broad scope and its surprising details, The WEIRDest People in the World explores how culture, institutions, and psychology shape one another, and explains what this means for both our most personal sense of who we are as individuals and also the large-scale social, political, and economic forces that drive human history. Includes black-and-white illustrations.

Nucleic Acid Transfection

Nucleic Acid Transfection PDF Author: Wolfgang Bielke
Publisher: Springer Science & Business Media
ISBN: 3642164293
Category : Science
Languages : en
Pages : 316

Book Description
Gene Delivery into Mammalian Cells: An Overview on Existing Approaches Employed In Vitro and In Vivo, by Peter Hahn and Elizabeth Scanlan * Strategies for the Preparation of Synthetic Transfection Vectors, by Asier Unciti-Broceta, Matthew N. Bacon, and Mark Bradley * Cationic Lipids: Molecular Structure/Transfection Activity Relationships and Interactions with Biomembranes, by Rumiana Koynova and Boris Tenchov * Hyperbranched Polyamines for Transfection, by Wiebke Fischer, Marcelo Calderon, and Rainer Haag * Carbohydrate Polymers for Nonviral Nucleic Acid Delivery, by Antons Sizovs, Patrick M. McLendon, Sathya Srinivasachari, and Theresa M. Reineke * Cationic Liposome–Nucleic Acid Complexes for Gene Delivery and Silencing: Pathways and Mechanisms for Plasmid DNA and siRNA, by Kai K. Ewert, Alexandra Zidovska, Ayesha Ahmad, Nathan F. Bouxsein, Heather M. Evans, Christopher S. McAllister, Charles E. Samuel, and Cyrus R. Safinya * Chemically Programmed Polymers for Targeted DNA and siRNA Transfection, by Eveline Edith Salcher and Ernst Wagner * Photochemical Internalization: A New Tool for Gene and Oligonucleotide Delivery, by Kristian Berg, Maria Berstad, Lina Prasmickaite, Anette Weyergang, Pål K. Selbo, Ida Hedfors, and Anders Høgset * Visualizing Uptake and Intracellular Trafficking of Gene Carriers by Single-Particle Tracking, by N. Ruthardt and C. Bräuchle

Natural Terpenoids as Messengers

Natural Terpenoids as Messengers PDF Author: Paul Harrewijn
Publisher: Springer Science & Business Media
ISBN: 9401007675
Category : Science
Languages : en
Pages : 445

Book Description
In contrast to books on specific bioactive compounds, this book deals with the role of mevalonic acid metabolites (isoprenoids and their derived structures) in metabolism, development, and functions of organisms, which, though diverse, show various levels of communication. Different disciplines are brought together in a discussion of the messenger functions of terpenoids within and between organisms belonging to five biological kingdoms: bacteria, fungi, plants, insects, and vertebrates (including humans). The present volume covers evolutionary aspects of terpenoids in plant physiology, plant-insect relationships, semiochemicals, and in life sciences, with special emphasis on cancer research and treatment. The book provides proposals for multidisciplinary model systems for the study of interrelationships of organisms utilizing terpenoid messengers, and discusses novel strategies for insect control and multifactor treatments of cancer utilizing terpenoids. This book is of interest to scientists, and students at an advanced level: biologists, plant breeders, pharmacologists, specialists in medical fields, especially oncologists, physicians in general, and anyone with a basic biochemical and physiological knowledge.

Bioanalytical Tools in Water Quality Assessment

Bioanalytical Tools in Water Quality Assessment PDF Author: Beate Escher
Publisher: IWA Publishing
ISBN: 1843393689
Category : Science
Languages : en
Pages : 273

Book Description
Part of Water Quality Set - Buy all four books and save over 30% on buying separately! Bioanalytical Tools in Water Quality Assessment reviews the application of bioanalytical tools to the assessment of water quality including surveillance monitoring. The types of water included range from wastewater to drinking water, including recycled water, as well as treatment processes and advanced water treatment. Bioanalytical Tools in Water Quality Assessment not only demonstrates applications but also fills in the background knowledge in toxicology/ecotoxicology needed to appreciate these applications. Each chapter summarises fundamental material in a targeted way so that information can be applied to better understand the use of bioanalytical tools in water quality assessment. Bioanalytical tools in Water Quality Assessment can be used by lecturers teaching academic and professional courses and also by risk assessors, regulators, experts, consultants, researchers and managers working in the water sector. It can also be a reference manual for environmental engineers, analytical chemists, and toxicologists. Authors: Beate Escher, National Research Centre for Environmental Toxicology (EnTox), The University of Queensland, Australia, Frederic Leusch, Smart Water Research Facility (G51), Griffith University Gold Coast Campus, Australia. With contributions by Heather Chapman and Anita Poulsen

Introduction to Single Cell Omics

Introduction to Single Cell Omics PDF Author: Xinghua Pan
Publisher: Frontiers Media SA
ISBN: 2889459209
Category :
Languages : en
Pages : 129

Book Description
Single-cell omics is a progressing frontier that stems from the sequencing of the human genome and the development of omics technologies, particularly genomics, transcriptomics, epigenomics and proteomics, but the sensitivity is now improved to single-cell level. The new generation of methodologies, especially the next generation sequencing (NGS) technology, plays a leading role in genomics related fields; however, the conventional techniques of omics require number of cells to be large, usually on the order of millions of cells, which is hardly accessible in some cases. More importantly, harnessing the power of omics technologies and applying those at the single-cell level are crucial since every cell is specific and unique, and almost every cell population in every systems, derived in either vivo or in vitro, is heterogeneous. Deciphering the heterogeneity of the cell population hence becomes critical for recognizing the mechanism and significance of the system. However, without an extensive examination of individual cells, a massive analysis of cell population would only give an average output of the cells, but neglect the differences among cells. Single-cell omics seeks to study a number of individual cells in parallel for their different dimensions of molecular profile on genome-wide scale, providing unprecedented resolution for the interpretation of both the structure and function of an organ, tissue or other system, as well as the interaction (and communication) and dynamics of single cells or subpopulations of cells and their lineages. Importantly single-cell omics enables the identification of a minor subpopulation of cells that may play a critical role in biological process over a dominant subpolulation such as a cancer and a developing organ. It provides an ultra-sensitive tool for us to clarify specific molecular mechanisms and pathways and reveal the nature of cell heterogeneity. Besides, it also empowers the clinical investigation of patients when facing a very low quantity of cell available for analysis, such as noninvasive cancer screening with circulating tumor cells (CTC), noninvasive prenatal diagnostics (NIPD) and preimplantation genetic test (PGT) for in vitro fertilization. Single-cell omics greatly promotes the understanding of life at a more fundamental level, bring vast applications in medicine. Accordingly, single-cell omics is also called as single-cell analysis or single-cell biology. Within only a couple of years, single-cell omics, especially transcriptomic sequencing (scRNA-seq), whole genome and exome sequencing (scWGS, scWES), has become robust and broadly accessible. Besides the existing technologies, recently, multiplexing barcode design and combinatorial indexing technology, in combination with microfluidic platform exampled by Drop-seq, or even being independent of microfluidic platform but using a regular PCR-plate, enable us a greater capacity of single cell analysis, switching from one single cell to thousands of single cells in a single test. The unique molecular identifiers (UMIs) allow the amplification bias among the original molecules to be corrected faithfully, resulting in a reliable quantitative measurement of omics in single cells. Of late, a variety of single-cell epigenomics analyses are becoming sophisticated, particularly single cell chromatin accessibility (scATAC-seq) and CpG methylation profiling (scBS-seq, scRRBS-seq). High resolution single molecular Fluorescence in situ hybridization (smFISH) and its revolutionary versions (ex. seqFISH, MERFISH, and so on), in addition to the spatial transcriptome sequencing, make the native relationship of the individual cells of a tissue to be in 3D or 4D format visually and quantitatively clarified. On the other hand, CRISPR/cas9 editing-based In vivo lineage tracing methods enable dynamic profile of a whole developmental process to be accurately displayed. Multi-omics analysis facilitates the study of multi-dimensional regulation and relationship of different elements of the central dogma in a single cell, as well as permitting a clear dissection of the complicated omics heterogeneity of a system. Last but not the least, the technology, biological noise, sequence dropout, and batch effect bring a huge challenge to the bioinformatics of single cell omics. While significant progress in the data analysis has been made since then, revolutionary theory and algorithm logics for single cell omics are expected. Indeed, single-cell analysis exert considerable impacts on the fields of biological studies, particularly cancers, neuron and neural system, stem cells, embryo development and immune system; other than that, it also tremendously motivates pharmaceutic RD, clinical diagnosis and monitoring, as well as precision medicine. This book hereby summarizes the recent developments and general considerations of single-cell analysis, with a detailed presentation on selected technologies and applications. Starting with the experimental design on single-cell omics, the book then emphasizes the consideration on heterogeneity of cancer and other systems. It also gives an introduction of the basic methods and key facts for bioinformatics analysis. Secondary, this book provides a summary of two types of popular technologies, the fundamental tools on single-cell isolation, and the developments of single cell multi-omics, followed by descriptions of FISH technologies, though other popular technologies are not covered here due to the fact that they are intensively described here and there recently. Finally, the book illustrates an elastomer-based integrated fluidic circuit that allows a connection between single cell functional studies combining stimulation, response, imaging and measurement, and corresponding single cell sequencing. This is a model system for single cell functional genomics. In addition, it reports a pipeline for single-cell proteomics with an analysis of the early development of Xenopus embryo, a single-cell qRT-PCR application that defined the subpopulations related to cell cycling, and a new method for synergistic assembly of single cell genome with sequencing of amplification product by phi29 DNA polymerase. Due to the tremendous progresses of single-cell omics in recent years, the topics covered here are incomplete, but each individual topic is excellently addressed, significantly interesting and beneficial to scientists working in or affiliated with this field.

Protein Purification

Protein Purification PDF Author: Jan-Christer Janson
Publisher: John Wiley & Sons
ISBN: 9780471186267
Category : Medical
Languages : en
Pages : 712

Book Description
This is a state-of-the-art sourcebook on modern high-resolution biochemical separation techniques for proteins. It contains all the basic theory and principles used in protein chromatography and electrophoresis.