Publications by authors named "Joanne Santini"

The new age of the phage.

Essays Biochem

December 2024

The discovery of viruses that can devour bacteria, bacteriophages (phages), was in 1915. Phages are ubiquitous, outnumbering the organisms they devour, and genomically, morphologically, and ecologically diverse. They were critical in our development of molecular biology and biotechnology tools and have been used as therapeutics for over 100 years, primarily in Eastern Europe with thousands of patients from all over the world treated in Georgia.

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Article Synopsis
  • Klebsiella pneumoniae is an opportunistic pathogen that poses significant health risks, and its treatment is complicated by the emergence of multidrug-resistant strains.
  • Bacteriophages that target K. pneumoniae produce specialized enzymes called depolymerases, which can break down the bacteria's protective capsules, making them potential new antimicrobials.
  • Understanding the structures and mechanisms of these depolymerases is crucial for developing effective therapies, as they show diversity in their function and specificity, suggesting potential for engineering improved enzymes for treatment.
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High-throughput sequencing for uncultivated viruses has accelerated the understanding of global viral diversity and uncovered viral genomes substantially larger than any that have so far been cultured. Notably, the Lak phages are an enigmatic group of viruses that present some of the largest known phage genomes identified in human and animal microbiomes, and are dissimilar to any cultivated viruses. Despite the wealth of viral diversity that exists within sequencing datasets, uncultivated viruses have rarely been used for taxonomic classification.

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Bioleaching offers a low-input method of extracting valuable metals from sulfide minerals, which works by exploiting the sulfur and iron metabolisms of microorganisms to break down the ore. Bioleaching microbes generate energy by oxidising iron and/or sulfur, consequently generating oxidants that attack sulfide mineral surfaces, releasing target metals. As sulfuric acid is generated during the process, bioleaching organisms are typically acidophiles, and indeed the technique is based on natural processes that occur at acid mine drainage sites.

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Arsenic contamination of groundwater is among one of the biggest health threats affecting millions of people in the world. There is an urgent need for efficient arsenic biosensors where the use of arsenic metabolizing enzymes can be explored. In this work, we have solved four crystal structures of arsenite oxidase (Aio) in complex with arsenic and antimony oxyanions and the structures determined correspond to intermediate states of the enzymatic mechanism.

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The arsenite oxidase (AioAB) from Pseudorhizobium banfieldiae sp. strain NT-26 catalyzes the oxidation of arsenite to arsenate and transfers electrons to its cognate electron acceptor cytochrome c (cytc). This activity underpins the ability of this organism to respire using arsenite present in contaminated environments.

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Arsenic is a ubiquitous toxic element, the global cycle of which is highly affected by microbial redox reactions and assimilation into organoarsenic compounds through sequential methylation reactions. While microbial biotransformation of arsenic has been studied for decades, the past years have seen the discovery of multiple new genes related to arsenic metabolism. Still, most studies focus on a small set of key genes or a small set of cultured microorganisms.

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  • Bacteriophages are viruses that rely on bacteria to reproduce but some have unique genetic codes that include reassigned stop codons, which are usually not compatible with bacterial translation systems.
  • A study of 9,422 phage genomes revealed that this stop-codon recoding occurs in various phage groups that infect bacteria found in human and animal gut microbiota, especially in genes related to phage structure and cell lysis.
  • The study suggests that these recoded stop codons may help delay the production of late-stage viral proteins, indicating that such changes can evolve quickly within closely related phage lineages.
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Lak megaphages are prevalent across diverse gut microbiomes and may potentially impact animal and human health through lysis of . Given their large genome size (up to 660 kbp), Lak megaphages are difficult to culture, and their identification relies on molecular techniques. Here, we present optimized protocols for identifying Lak phages in various microbiome samples, including procedures for DNA extraction, followed by detection and quantification of genes encoding Lak structural proteins using diagnostic endpoint and SYBR green-based quantitative PCR, respectively.

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Lak phages with alternatively coded ∼540 kbp genomes were recently reported to replicate in in microbiomes of humans that consume a non-Western diet, baboons, and pigs. Here, we explore Lak phage diversity and broader distribution using diagnostic polymerase chain reaction and genome-resolved metagenomics. Lak phages were detected in 13 animal types, including reptiles, and are particularly prevalent in pigs.

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The anaerobic bacterium respires using the oxyanion arsenate (AsO) as the terminal electron acceptor, where it is reduced to arsenite (AsO) while concomitantly oxidizing various organic (e.g., acetate) electron donors.

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The family Rhizobiaceae includes many genera of soil bacteria, often isolated for their association with plants. Herein, we investigate the genomic diversity of a group of Rhizobium species and unclassified strains isolated from atypical environments, including seawater, rock matrix or polluted soil. Based on whole-genome similarity and core genome phylogeny, we show that this group corresponds to the genus Pseudorhizobium.

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Many bacteria can exchange genetic material through horizontal gene transfer (HGT) mediated by plasmids and plasmid-borne transposable elements. Here, we study the population structure and dynamics of over 10,000 bacterial plasmids, by quantifying their genetic similarities and reconstructing a network based on their shared k-mer content. We use a community detection algorithm to assign plasmids into cliques, which correlate with plasmid gene content, bacterial host range, GC content, and existing classifications based on replicon and mobility (MOB) types.

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Article Synopsis
  • - Researchers discovered hundreds of bacteriophage genomes over 200 kilobases, including the largest known at 735 kb, showing that phages can have significantly larger genetic material than previously thought.
  • - Many of these phages possess unique genetic elements, such as previously unidentified CRISPR-Cas systems and various tRNA-related genes, hinting at complex interactions with their bacterial hosts.
  • - The study classifies major groups of these large phages from various ecosystems around the world, suggesting they play a key role in microbial interactions and could influence microbial diversity across different environments.
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The original version of this Article contained errors in Fig. 4. In panel a, the labels 'F420-reducing NiFe hydrogenase (group 3a)' and 'Group 2 NiFe hydrogenase' were misplaced.

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Bacteriophages (phages) dramatically shape microbial community composition, redistribute nutrients via host lysis and drive evolution through horizontal gene transfer. Despite their importance, much remains to be learned about phages in the human microbiome. We investigated the gut microbiomes of humans from Bangladesh and Tanzania, two African baboon social groups and Danish pigs; many of these microbiomes contain phages belonging to a clade with genomes >540 kilobases in length, the largest yet reported in the human microbiome and close to the maximum size ever reported for phages.

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The evolution of aerobic respiration was likely linked to the origins of oxygenic Cyanobacteria. Close phylogenetic neighbors to Cyanobacteria, such as Margulisbacteria (RBX-1 and ZB3), Saganbacteria (WOR-1), Melainabacteria and Sericytochromatia, may constrain the metabolic platform in which aerobic respiration arose. Here, we analyze genomic sequences and predict that sediment-associated Margulisbacteria have a fermentation-based metabolism featuring a variety of hydrogenases, a streamlined nitrogenase, and electron bifurcating complexes involved in cycling of reducing equivalents.

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Arsenate respiration by bacteria was discovered over two decades ago and is catalyzed by diverse organisms using the well-conserved Arr enzyme complex. Until now, the mechanisms underpinning this metabolism have been relatively opaque. Here, we report the structure of an Arr complex (solved by X-ray crystallography to 1.

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The majority of the population of Bangladesh (90%) rely on untreated groundwater for drinking and domestic use. At the point of collection, 40% of these supplies are contaminated with faecal indicator bacteria (FIB). Recent studies have disproved the theory that latrines discharging to shallow aquifers are the major contributor to this contamination.

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Article Synopsis
  • Over 140 million people globally are affected by arsenic contamination in drinking water, which poses significant toxicity to humans.
  • Arsenite and arsenate, inorganic forms of arsenic, can be used by some microbes as energy sources, highlighting a unique aspect of microbial respiration.
  • The study reveals AioX and its related proteins function as key regulators in arsenic metabolism, with structural analysis showing specific adaptations in protein binding that influence selectivity between arsenite and arsenate, indicating a complex evolutionary relationship.
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Rhizobium sp. str. NT-26 is a Gram-negative facultative chemolithoautotrophic arsenite oxidizer that has been used as a model organism to study various aspects of arsenite oxidation including the regulation of arsenite oxidation.

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Arsenic is a widely distributed environmental toxin whose presence in drinking water poses a threat to >140 million people worldwide. The respiratory enzyme arsenite oxidase from various bacteria catalyses the oxidation of arsenite to arsenate and is being developed as a biosensor for arsenite. The arsenite oxidase from Rhizobium sp.

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