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Single-molecule Dynamics in Protein Interactions: Characterization of RarA and RecD2 of Bacillus subtilis

Maintenance of genome integrity is one of the crucial functions in life, to preserve the appropriate genetic information, being homologous recombination a key process in the DNA repair. I have used a novel technique, using slim-field microscopy to obtain single-molecule dynamics of two poorly descri...

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Bibliographische Detailangaben
1. Verfasser: Romero Gonzalez, Hector
Beteiligte: Graumann, Peter L. (Prof. Dr.) (BetreuerIn (Doktorarbeit))
Format: Dissertation
Sprache:Englisch
Veröffentlicht: Philipps-Universität Marburg 2018
Schlagworte:
fluorescence microscopy
Mikroskopische Technik
Chemie
DNA Reparatur
single-molecule microscopy
homologe Rekombination
Molekulargenetik
Bacillus subtilis
Fluoreszenzmikroskopie
DNA repair
Bakterien
Homologous recombination
Chemistry & allied sciences
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Zusammenfassung:Maintenance of genome integrity is one of the crucial functions in life, to preserve the appropriate genetic information, being homologous recombination a key process in the DNA repair. I have used a novel technique, using slim-field microscopy to obtain single-molecule dynamics of two poorly described proteins, RarA and RecD2, in different recombination deficient mutants and conditions to characterize them. Single-molecule microscopy has been shown as a powerful method for in vivo characterization of proteins and its interactions. Together with genetics, I have added a new level of complexity in the regulation of homologous recombination as a multiway process in which many factors are involved in different avenues with partially overlapping functions depending on the kind of DNA damage generated. I have characterized RarA and RecD2 as factors involved in recombination, but also in replication of the DNA, being part of both RecA-independent and RecA-dependent replication progression, and antagonistic regulators of RecA filamentation. RarA plays its role in replication through interactions with DnaB, and in recombination as a RecA positive regulator through its interactions with RecA, RecO, RecR, RecD2 and RecU. RarA is regulated by the RecQ-like helicases RecQ and RecS. RecD2 plays a role in chromosomal segregation that becomes essential in the absence of RecG or RuvAB, and is a negative regulator in homologous recombination that interacts with RecA, RarA, RecX, RecF and PcrA.
Umfang:105 Seiten
DOI:10.17192/z2018.0110

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