Publications by authors named "Prune Leroy"

Soil contaminants may restrict soil functions. A promising soil remediation method is amendment with biochar, which has the potential to both adsorb contaminants and improve soil health. However, effects of biochar amendment on soil-plant nitrogen (N) dynamics and N cycling microbial guilds in contaminated soils are still poorly understood.

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Homologous recombination is essential for the accurate repair of double-stranded DNA breaks (DSBs). Initially, the RecBCD complex resects the ends of the DSB into 3' single-stranded DNA on which a RecA filament assembles. Next, the filament locates the homologous repair template on the sister chromosome.

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How fast can a cell locate a specific chromosomal DNA sequence specified by a single-stranded oligonucleotide? To address this question, we investigate the intracellular search processes of the Cas9 protein, which can be programmed by a guide RNA to bind essentially any DNA sequence. This targeting flexibility requires Cas9 to unwind the DNA double helix to test for correct base pairing to the guide RNA. Here we study the search mechanisms of the catalytically inactive Cas9 (dCas9) in living by combining single-molecule fluorescence microscopy and bulk restriction-protection assays.

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Transcription factors mediate gene regulation by site-specific binding to chromosomal operators. It is commonly assumed that the level of repression is determined solely by the equilibrium binding of a repressor to its operator. However, this assumption has not been possible to test in living cells.

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Article Synopsis
  • Transcription factors (TFs) are proteins that control gene expression by binding to specific sites on chromosomes, using a combination of 1D sliding on DNA and 3D diffusion in the cytoplasm to locate these sites quickly.
  • A new single-molecule assay was developed to study the sliding behavior of the lac repressor in living bacteria, revealing that it can slide approximately 45 base pairs of DNA and may encounter obstacles from other DNA-bound proteins.
  • The study found that the lac repressor often slides over its target site multiple times before binding, indicating a balance between searching quickly through nonspecific regions and making fast, effective connections at specific sequences.
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We have shown previously that lack of molybdenum cofactor (MoCo) in Escherichia coli leads to hypersensitivity to the mutagenic and toxic effects of N-hydroxylated base analogues, such as 6-N-hydroxylaminopurine (HAP). However, the nature of the MoCo-dependent mechanism is unknown, as inactivation of all known and putative E. coli molybdoenzymes does not produce any sensitivity.

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The translational initiation region (TIR) of the Escherichia coli rpsA gene, which encodes ribosomal protein S1, shows a number of unusual features. It extends far upstream (to position -91) of the initiator AUG, it lacks a canonical Shine-Dalgarno sequence (SD) element, and it can fold into three successive hairpins (I, II, and III) that are essential for high translational activity. Two conserved GGA trinucleotides, present in the loops of hairpins I and II, have been proposed to form a discontinuous SD.

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