Biochemical and Genetic Characterization of the E. Coli RadD and RecG Proteins PDF Download
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Author: Nina Bonde
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
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Faithful replication of the genomic material is a requirement for successful cell propagation and division in all organisms. In Escherichia coli, the RadD and RecG proteins play important roles in DNA repair and recombination. This thesis reveals an important molecular function for RadD and examines RecG protein and gene interactions that help modulate and define its function in cells. We describe a new molecular function for RadD. RadD accelerates RecA-mediated strand exchange in vitro. The ability of RadD to accelerate strand exchange is dependent on both its ATP hydrolysis activity and the presence of RecA. RadD is unable to remodel recombination intermediate molecules in the absence of RecA. The dependence on RecA to carry out this function places RadD in a new class of enzymes. RecG physically interacts with the single-stranded DNA-binding protein (SSB). We report that this interaction is mediated by the highly conserved SSB C-terminal tip (SSB-Ct). The SSB-Ct docks onto a conserved region of the RecG Helicase Domain 2, and the SSB-Ct binding pocket on RecG is framed by three Arg residues. Charge-reversal mutations to any of the three Arg residues results in loss of the RecG-SSB interaction and cellular defects, including induced SOS response and sensitivity to ultraviolet radiation. As a complement to this study, we also report that the intrinsically disorder linker of SSB is not necessary under normal growth conditions and likely acts as a simple spacer between the SSB OB domain and SSB-Ct interaction motif. RecG function has been difficult to elucidate in vivo. We identify recG genetic interactions by transposon sequencing to gain insight into the activity of recG in cells. The strongest genetic interactions occur with the dam, uvrD, rnhA, radA, and rep genes. Viability of strains lacking recG in addition to one of these five genes is generally improved by additional loss of the recF or recO genes. recG mutants that are deficient in SSB binding produce similar phenotypes as cells lacking recG. Collectively, these results support a role for RecG in alleviating toxic recombination intermediates.
Author: Nina Bonde
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
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Book Description
Faithful replication of the genomic material is a requirement for successful cell propagation and division in all organisms. In Escherichia coli, the RadD and RecG proteins play important roles in DNA repair and recombination. This thesis reveals an important molecular function for RadD and examines RecG protein and gene interactions that help modulate and define its function in cells. We describe a new molecular function for RadD. RadD accelerates RecA-mediated strand exchange in vitro. The ability of RadD to accelerate strand exchange is dependent on both its ATP hydrolysis activity and the presence of RecA. RadD is unable to remodel recombination intermediate molecules in the absence of RecA. The dependence on RecA to carry out this function places RadD in a new class of enzymes. RecG physically interacts with the single-stranded DNA-binding protein (SSB). We report that this interaction is mediated by the highly conserved SSB C-terminal tip (SSB-Ct). The SSB-Ct docks onto a conserved region of the RecG Helicase Domain 2, and the SSB-Ct binding pocket on RecG is framed by three Arg residues. Charge-reversal mutations to any of the three Arg residues results in loss of the RecG-SSB interaction and cellular defects, including induced SOS response and sensitivity to ultraviolet radiation. As a complement to this study, we also report that the intrinsically disorder linker of SSB is not necessary under normal growth conditions and likely acts as a simple spacer between the SSB OB domain and SSB-Ct interaction motif. RecG function has been difficult to elucidate in vivo. We identify recG genetic interactions by transposon sequencing to gain insight into the activity of recG in cells. The strongest genetic interactions occur with the dam, uvrD, rnhA, radA, and rep genes. Viability of strains lacking recG in addition to one of these five genes is generally improved by additional loss of the recF or recO genes. recG mutants that are deficient in SSB binding produce similar phenotypes as cells lacking recG. Collectively, these results support a role for RecG in alleviating toxic recombination intermediates.
Author: Zachary Joseph Romero
Publisher:
ISBN:
Category :
Languages : en
Pages : 214
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Book Description
In every round of replication, the replisome encounters various forms of damage such as DNA breaks, protein-DNA crosslinks, and oxidized bases. Escherichia coli has evolved multiple pathways to repair the various types of DNA damage that accumulates in the cell. Recombinational repair is a non-mutagenic repair pathway that utilizes a neighboring undamaged duplex to restore lost genetic information. Our lab has recently implicated the E. coli Uup and RadD in the recombinational repair of a wide variety of DNA lesions. We show that the SF2 helicase RadD is essential when another helicase, RecG, is removed from the cell. This synthetically lethal phenotype allowed us to leverage existing knowledge of RecG to better understand the functions that become essential in its absence. Functions that are specifically carried out by RadD. This synthetically lethal phenotype is rapidly suppressed by mutations that affect replication restart (PriA) and homologous recombination (RecA). We also identify that a full gene deletion of uup is capable of suppressing the [Delta]rad[Delta]recG phenotype. By using a variety of biochemical and genetic approaches we show that Uup and RadD are involved in the processing of the intermediates generated by RecA-dependent and independent recombination in every replication cycle. When this activity is lost, these intermediates are targeted for alternative processing, by PriA and Uup. This alternative processing has deleterious consequences and results in cell death. Altogether, the results presented here shed further light on Uup and RadD and their roles in recombinational repair.
Author: Yuejin Hua
Publisher: Frontiers Media SA
ISBN: 288974194X
Category : Science
Languages : en
Pages : 154
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Publisher:
ISBN:
Category : Dissertations, Academic
Languages : en
Pages : 780
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ISBN:
Category : Medicine
Languages : en
Pages : 1320
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Author: Andrei Kuzminov
Publisher: Landes Bioscience
ISBN:
Category : Medical
Languages : en
Pages : 234
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Author: Steffen Backert
Publisher: Springer Nature
ISBN: 3030654818
Category : Medical
Languages : en
Pages : 334
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This edited volume explores Campylobacter species, which are some of the most important foodborne pathogens. Above all, contaminated poultry meat can cause human gastroenteritis in both developed and developing countries. The respective contributions reveal how these infections can also increase the risk of generalized paralytic diseases such as Guillain-Barré syndrome, Miller-Fisher syndrome, and Chinese paralytic syndrome. Due to their influence on the nervous system, circulatory system, and various organs, Campylobacter infections represent a serious public health concern. Campylobacter can be effectively combated by addressing the hygienic conditions in both food production and human lifestyles. Accordingly, the authors put forward a One Health perspective, which provides readers with essential insights into the basic biology of Campylobacter, as well as practical guidance on aspects ranging from food production to the clinical treatment of infections. Chapters 'Population Biology and Comparative Genomics of Campylobacter Species' and 'Natural Competence and Horizontal Gene Transfer in Campylobacter' are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
Author: Mousumi Debnath
Publisher: Springer Science & Business Media
ISBN: 9048132614
Category : Medical
Languages : en
Pages : 527
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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.
Author: Paul James Smith
Publisher: Oxford University Press, USA
ISBN:
Category : Medical
Languages : en
Pages : 260
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Book Description
The processes of DNA recombination and repair are vital to cell integrity - an error can lead to disease such as cancer. It is therefore a large and exciting area of research and is also taught on postgraduate and undergraduate courses. This book is not a comprehensive view of the field, but a selection of the issues currently at the forefront of knowledge.
Author: Behzad Rad
Publisher:
ISBN:
Category :
Languages : en
Pages : 398
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