Publications by authors named "Micaela Cerletti"

Inducible promoters are one of cellular and molecular biology's most important technical tools. The ability to deplete, replete, and overexpress genes on demand is the foundation of most functional studies. Here, we developed and characterized a new xylose-responsive promoter (Pxyl), the second inducible promoter system for the model haloarcheon .

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Phytoene synthase (PSY) converts two molecules of geranyl-geranyl diphosphate to phytoene, the key regulatory step in carotenogenesis. However, post-translational mechanisms that control PSY expression are scarcely understood. Carotenoid biosynthesis (mainly bacterioruberin) is a distinctive feature of haloarchaea thriving in hypersaline environments.

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Carotenoids are bioactive molecules known to promote human health. Many extreme halophilic archaea synthesize carotenoids, mainly represented by C bacterioruberin (BR) and its derivatives. BR has a potent antioxidant capacity, even higher than that of β-carotene, thus, there is an increasing interest to advance the study of its biological properties as well as to extend its current applications.

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Article Synopsis
  • Coimmunoprecipitation is a key technique for studying protein-protein interactions in living organisms, and this study focuses on optimizing it for the haloarchaeon Haloferax volcanii to explore relationships involving the LonB protease.* -
  • The adapted protocol involves in vivo cross-linking of proteins using either dithiobis(succinimidyl propionate) or formaldehyde, and then isolating the proteins with anti-LonB antibodies attached to Protein A-Sepharose beads.* -
  • After isolating the protein interactions, tryptic digestion and Mass Spectrometry are used to analyze the peptides, helping to identify and quantify the proteins that interact with LonB.*
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The cellular protein repertoire is highly dynamic and responsive to internal or external stimuli. Its changes are largely the consequence of the combination of protein synthesis and degradation, referred collectively as protein turnover. Different proteomics techniques have been developed to determine the whole proteome turnover of a cell, but very few have been applied to archaea.

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Article Synopsis
  • Proteolysis is crucial for various cellular processes like protein quality control, signaling, and cell envelope formation.
  • Archaea, a diverse group of prokaryotes, thrive in extreme environments, offering insights into proteases' roles in cell physiology and adaptation.
  • This review highlights recent progress on archaeal membrane proteases, particularly their biological functions and potential targets, focusing on a specific model organism known as haloarchaeon.
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  • Sperm capacitation is essential for fertilization in mammals, and decapacitation factors (DFs) like SPINK3 inhibit premature capacitation.
  • SPINK3 binds to the sperm membrane through specific lipid interactions, particularly in lipid rafts, and its binding can be blocked by cholera toxin subunit B (CTB).
  • During the capacitation process, SPINK3 is gradually removed from sperm in the female reproductive tract, starting from the sperm's head and then from its tail, correlating with the presence of acrosome-reacted sperm when SPINK3 is absent.
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While many aspects of archaeal cell biology remain relatively unexplored, systems biology approaches like mass spectrometry (MS) based proteomics offer an opportunity for rapid advances. Unfortunately, the enormous amount of MS data generated often remains incompletely analyzed due to a lack of sophisticated bioinformatic tools and field-specific biological expertise for data interpretation. Here we present the initiation of the Archaeal Proteome Project (ArcPP), a community-based effort to comprehensively analyze archaeal proteomes.

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The dynamic changes that take place along the phases of microbial growth (lag, exponential, stationary, and death) have been widely studied in bacteria at the molecular and cellular levels, but little is known for archaea. In this study, a high-throughput approach was used to analyze and compare the proteomes of two haloarchaea during exponential and stationary growth: the neutrophilic Haloferax volcanii and the alkaliphilic Natrialba magadii. Almost 2000 proteins were identified in each species (≈50% of the predicted proteome).

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The membrane protease LonB is an essential protein in the archaeon Haloferax volcanii and globally impacts its physiology. However, natural substrates of the archaeal Lon protease have not been identified. The whole proteome turnover was examined in a H.

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Article Synopsis
  • - Rhomboids are intramembrane serine proteases crucial for cell signaling, but their role in prokaryotes, especially Archaea, is not well understood.
  • - A study on a rhomboid homologue deletion mutant (ΔrhoII) in Haloferax volcanii revealed issues like reduced motility and N-glycosylation defects, prompting an analysis of protein changes in the mutant compared to the wild-type strain.
  • - Out of 1847 proteins identified, 103 showed differences in quantity and 99 had altered processing, highlighting key proteins linked to motility and glycosylation issues in the mutant, and marking the first investigation of a rhomboid protease's impact
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This data article provides information in support of the research article "Global role of the membrane protease LonB in Archaea: Potential protease targets revealed by quantitative proteome analysis of a lonB mutant in Haloferax volcanii" [1]. The proteome composition of a wt and a LonB protease mutant strain (suboptimal expression) in the archaeon Haloferax volcanii was assessed by a quantitative shotgun proteomic approach. Membrane and cytosol fractions of H.

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Unlabelled: The membrane-associated LonB protease is essential for viability in Haloferax volcanii, however, the cellular processes affected by this protease in archaea are unknown. In this study, the impact of a lon conditional mutation (down-regulation) on H. volcanii physiology was examined by comparing proteomes of parental and mutant cells using shotgun proteomics.

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The function of membrane proteases range from general house-keeping to regulation of cellular processes. Although the biological role of these enzymes in archaea is poorly understood, some of them are implicated in the biogenesis of the archaeal cell envelope and surface structures. The membrane-bound ATP-dependent Lon protease is essential for cell viability and affects membrane carotenoid content in Haloferax volcanii.

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Although homologs of the ATP-dependent Lon protease exist in all domains of life, the relevance of this protease in archaeal physiology remains a mystery. In this study, we have constructed and phenotypically characterized deletion and conditional lon mutants in the model haloarchaeon Haloferax volcanii to elucidate the role of the unusual membrane-bound LonB protease in archaea. Hvlon could be deleted from the chromosome only when a copy of the wild type gene was provided in trans suggesting that Lon is essential for survival in this archaeon.

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