Research, Research Reports

Detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its first variants in fourplex real-time quantitative reverse transcription-PCR assays

Mathieu Durand1, Philippe Thibault1, Simon Lévesque2,3, Ariane Brault4, Alex Carignan2, Louis Valiquette2, Philippe Martin2 and Simon Labbé4

The early diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections is required to identify and isolate contagious patients to prevent further transmission of SARS-CoV-2. In this study, we present a multitarget real-time TaqMan reverse transcription PCR (rRT-PCR) assay for the quantitative detection of SARS-CoV-2 and some of its circulating variants harboring mutations that give the virus a selective advantage. Seven different primer-probe sets that included probes containing locked nucleic acid (LNA) nucleotides were designed to amplify specific wild-type and mutant sequences in Orf1ab, Envelope (E), Spike (S), and Nucleocapsid (N) genes (...)

Forced association of SARS-CoV-2 proteins with the yeast proteome perturb vesicle trafficking

Cinzia Klemm1,#, Henry Wood1,#, Grace Heredge Thomas1,#, Guðjón Ólafsson1,2, Mara Teixeira Torres1 and Peter H. Thorpe1

This work demonstrates that the yeast Synthetic Physical Interactions method is a rapid way to identify potential functions of ectopic viral proteins.

Airborne bacteria in show caves from Southern Spain

Irene Dominguez-Moñino1, Valme Jurado1, Miguel Angel Rogerio-Candelera1, Bernardo Hermosin1 and Cesareo Saiz-Jimenez1

This study analyzes the factors conditioning the diversity of airborne bacteria recorded in three Andalusian show caves, subjected to different managements.

Landscapes and bacterial signatures of mucosa-associated intestinal microbiota in Chilean and Spanish patients with inflammatory bowel disease

Nayaret Chamorro1,#, David A. Montero1,2,#, Pablo Gallardo3, Mauricio Farfán3, Mauricio Contreras4, Marjorie De la Fuente2, Karen Dubois2, Marcela A. Hermoso2, Rodrigo Quera5,6, Marjorie Pizarro-Guajardo7,8,9, Daniel Paredes-Sabja7,8,9, Daniel Ginard10, Ramon Rosselló-Móra11 and Roberto Vidal1,8,12

This study investigates the landscapes and alterations of mucosa-associated intestinal microbiota in patients with inflammatory bowel diseases, which cause chronic inflammation of the gut, including ulcerative colitis and Crohn’s disease.

Genome, transcriptome and secretome analyses of the antagonistic, yeast-like fungus Aureobasidium pullulans to identify potential biocontrol genes

Maria Paula Rueda-Mejia1, Lukas Nägeli1, Stefanie Lutz2, Richard D. Hayes3, Adithi R. Varadarajan2, Igor V. Grigoriev3,4, Christian H. Ahrens2,5 and Florian M. Freimoser1

This study highlights the value of a sequential approach starting with genome mining and consecutive transcriptome and secretome analyses in order to identify a limited number of potential target genes for detailed, functional analyses in Aureobasidium pullulans.

Barcode sequencing and a high-throughput assay for chronological lifespan uncover ageing-associated genes in fission yeast

Catalina A. Romila1,#, StJohn Townsend1,2,#, Michal Malecki1,3, Stephan Kamrad1,2,4, María Rodríguez-López1, Olivia Hillson1, Cristina Cotobal1, Markus Ralser2,5 and Jürg Bähler1

This work presents two approaches to study chronological lifespan (CLS) for medium- to high-throughput applications, a method for Bar-seq to identify mutants showing altered CLS and a novel medium-throughput colony-forming units assay that can be largely automated by robotics.

Proanthocyanidin-enriched cranberry extract induces resilient bacterial community dynamics in a gnotobiotic mouse model

Catherine C. Neto1,2,#, Benedikt M. Mortzfeld3,#, John R. Turbitt1,2, Shakti K. Bhattarai3, Vladimir Yeliseyev4, Nicholas DiBenedetto4, Lynn Bry4 and Vanni Bucci2,3

This study investigates the effect of a water-soluble, proanthocyanidin-rich cranberry juice extract on the short-term dynamics of a human-derived bacterial community in a gnotobiotic mouse model.

Dry biocleaning of artwork: an innovative methodology for Cultural Heritage recovery?

Giancarlo Ranalli1, Pilar Bosch-Roig2, Simone Crudele1, Laura Rampazzi3,4, Cristina Corti3 and Elisabetta Zanardini5

This work proposes an innovative methodology based on applied biotechnology for the recovery of altered stonework: the "dry biocleaning", which envisages the use of dehydrated microbial cells without the use of free water or gel-based matrices.

Aeration mitigates endoplasmic reticulum stress in Saccharomyces cerevisiae even without mitochondrial respiration

Huong Thi Phuong1, Yuki Ishiwata-Kimata1, Yuki Nishi1, Norie Oguchi1, Hiroshi Takagi1 and Yukio Kimata1

This work demonstrates a scenario, in which aeration acts beneficially on Saccharmyces cerevisiae cells even under fermentative conditions.

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Filamentation protects Candida albicans from amphotericin B-induced programmed cell death via a mechanism involving the yeast metacaspase, MCA1

April 25, 2016

Candida albicans proliferates in two distinct cell types: blastopores and filaments. Programmed cell death is a controlled form of cell suicide that occurs when C. albicans cells are exposed to fungicidal drugs like amphotericin B and caspofungin, and to other stressful conditions. We provide evidence that programmed cell death is cell-type specific in yeast: Filamentous C. albicans cells are more resistant to amphotericin B- and caspofungin-induced programmed cell death than their blastospore counterparts. Our genetic data suggest that this phenomenon is mediated by a protective mechanism involving the yeast metacaspase, MCA1.

Formaldehyde fixation is detrimental to actin cables in glucose-depleted S. cerevisiae cells

April 13, 2016

Actin filaments form cortical patches and emanating cables in fermenting cells of Saccharomyces cerevisiae. We assume that stability of actin cables reflects the metabolic status of the cell. Based on comparison of live and formaldehyde-fixed cells, our data suggest that formaldehyde affects respiration before fixation and this uneven signaling results in destabilization of actin cables in glucose-deprived cells.

Insights into dynamin-associated disorders through analysis of equivalent mutations in the yeast dynamin Vps1

March 22, 2016

The dynamins represent a superfamily of proteins that have been shown to function in a wide range of membrane fusion and fission events. An increasing number of mutations in the human classical dynamins, Dyn-1 and Dyn-2 has been reported, with diseases caused by these changes ranging from Charcot-Marie-Tooth disorder to epileptic encephalopathies. This study aimed to use the dynamin-like protein Vps1 of Saccharomyces cerevisiae as a model to gain insights into the mechanistic defects caused by specific dynamin mutations considered to underlie a number of diseases.

Genomic saturation mutagenesis and polygenic analysis identify novel yeast genes affecting ethyl acetate production, a non-selectable polygenic trait

March 18, 2016

Isolation of mutants in populations of microorganisms has been a valuable tool in experimental genetics for decades. The main disadvantage, however, is the inability of isolating mutants in non-selectable polygenic traits. Our study shows that genomic saturation mutagenesis combined with complex trait polygenic analysis could be used successfully to identify causative alleles underlying many non-selectable, polygenic traits in small collections of haploid strains with multiple induced mutations.

Differentiated cytoplasmic granule formation in quiescent and non-quiescent cells upon chronological aging

March 3, 2016

Stationary phase cultures represent a complicated cell population comprising at least two different cell types, quiescent (Q) and non-quiescent (NQ) cells. The authors show that the cell fate of NQ cells is largely irreversible even if they are allowed to reenter mitosis. Their results reveal that the formation of different granule structures may represent the early stage of cell type differentiation in yeast stationary phase cultures.

Towards understanding the gliotoxin detoxification mechanism: in vivo thiomethylation protects yeast from gliotoxin cytotoxicity

February 19, 2016

Gliotoxin is a mycotoxin produced by some species of ascomycete fungi including the opportunistic human pathogen Aspergillus fumigatus. In order to produce gliotoxin the host organism needs to have evolved a self-protection mechanism. The authors demonstrate that the activity of a novel thiomethyltransferase is requiered for protection against exogenous gliotoxin and provide implications for understanding the evolution of gliotoxin self-protection mechanisms.

Mitochondrial proteomics of the acetic acid – induced programmed cell death response in a highly tolerant Zygosaccharomyces bailii – derived hybrid strain

January 22, 2016

Very high concentrations of acetic acid at low pH induce programmed cell death (PCD) in both the experimental model Saccharomyces cerevisiae and in Zygosaccharomyces bailii, the latter being considered the most problematic acidic food spoilage yeast due to its remarkable intrinsic resistance to this food preservative. This study offers insights into the mechanisms involved in acetic acid - induced PCD in the Z. bailii-derived hybrid strain ISA1307 by analyzing the yeast mitochondrial protein expression profile of cells challenged by acetic acid.

The transcriptional repressor Sum1p counteracts Sir2p in regulation of the actin cytoskeleton, mitochondrial quality control and replicative lifespan in Saccharomyces cerevisiae

January 18, 2016

Increasing the stability or dynamics of the actin cytoskeleton can extend lifespan in C. elegans and S. cerevisiae. Actin cables of budding yeast, bundles of actin filaments that mediate cargo transport, affect lifespan control through effects on mitochondrial quality control. Here, we report that Sum1p and Sir2p inversely regulate actin and mitochondrial maintenance, as well as lifespan.

Inhibition of Aβ42 oligomerization in yeast by a PICALM ortholog and certain FDA approved drugs

January 18, 2016

The formation of small Aβ42 oligomers has been implicated as a toxic species in Alzheimer disease (AD). Here, we show that the mechanism of the PICALM, human AD risk factor, is likely to reduce the level of Aβ42 oligomers in cells. We screened FDA-approved drugs to identify candidates that prevent the formation of Aβ42 small oligomers using the yeast Aβ42-RF reporter system. We also showed that each of the drug hits counteract yeast and mammalian cell toxicity associated with Aβ42 small aggregates.

Global translational impacts of the loss of the tRNA modification t6A in yeast

December 18, 2015

The universal tRNA modification t6A is found at position 37 of nearly all tRNAs decoding ANN codons. Analysis of codon occupancy rates suggests that one of the major roles of t6A is to homogenize the process of elongation by slowing the elongation rate at codons decoded by high abundance tRNAs and I34:C3 pairs while increasing the elongation rate of rare tRNAs and G34:U3 pairs. This work reveals that the consequences of t6A absence are complex and multilayered and has set the stage to elucidate the molecular basis of the observed phenotypes.

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