Research, Research Reports
Expansion of metabolically labelled endocytic organelles and cytoskeletal cell structures in Giardia lamblia using optimised U- ExM protocols
Clirim Jetishi1,2,a, Erina A. Balmer1,2,a, Bianca M. Berger1,2,a, Carmen Faso1,3,4 and Torsten Ochsenreiter1
Understanding cellular ultrastructure is tightly bound to microscopic resolution and the ability to identify individual components at that resolution. In this study we demonstrate mostly isotropic 4.5-fold expansion of several different compartments in Giardia cells and present an optimised, shortened, and modular protocol that can be swiftly adjusted to the investigators needs.
Polyadenylated versions of small non-coding RNAs in Saccharomyces cerevisiae are degraded by Rrp6p/Rrp47p independent of the core nuclear exosome
Anusha Chaudhuri1,#, Soumita Paul2,#, Mayukh Banerjea2 and Biswadip Das2
In this investigation, we unveiled a novel functional role of the major nuclear 3′→5′ exoribonuclease, Rrp6p, and its cofactor Rrp47p in the degradation of polyadenylated versions of several mature sncRNAs, including 5S, 5.8S rRNAs, all sn- and some select snoRNAs in the baker’s yeast S. cerevisiae.
Exploring carbon source related localization and phosphorylation in the Snf1/Mig1 network using population and single cell-based approaches
Svenja Braam1, Farida Tripodi2, Linnea Österberg1,3, Sebastian Persson1, Niek Welkenhuysen1, Paola Coccetti2 and Marija Cvijovic1
In this work we set out to explore the relationship between the subcellular localization and regulation of kinases in the context of carbon source signaling. The data presented in this paper reinforce the notion that not only the activation/inactivation of kinases but also their subcellular localization and that of their targets influence fate decisions in response to environmental changes.
A Modular Cloning Toolkit for the production of recombinant proteins in Leishmania tarentolae
Katrin Hieronimus1,2,#, Tabea Donauer1,2,#, Jonas Klein1,#, Bastian Hinkel1,#, Julia Vanessa Spänle1,#, Anna Probst1,#, Justus Niemeyer1,#, Salina Kibrom1, Anna Maria Kiefer1, Luzia Schneider2, Britta Husemann2, Eileen Bischoff2, Sophie Möhring2, Nicolas Bayer1, Dorothée Klein1, Adrian Engels1, Benjamin Gustav Ziehmer2, Julian Stieß3, Pavlo Moroka1, Michael Schroda1, and Marcel Deponte2
Modular Cloning (MoClo) is based on libraries of standardized genetic parts that can be directionally assembled via Golden Gate cloning in one-pot reactions into transcription units and multigene constructs. We established a MoClo toolkit and exemplified its application for the production of recombinant proteins in L. tarentolae.
A metagenomic approach to unveil the association between fecal gut microbiota and short-chain fatty acids in diarrhea caused by diarrheagenic Escherichia coli in children
Pablo Gallardo1,2, Mariana Izquierdo2, Tomeu Viver3, Esteban Bustos-Caparros3, Dana Piras2, Roberto M. Vidal4, Hermie J.M. Harmsen1 and Mauricio J. Farfan2
Diarrheagenic Escherichia coli (DEC) is the main cause of diarrhea in children under five years old. The virulence of DEC is tightly regulated by environmental signals influenced by the gut microbiota and its metabolites. Our results increase the knowledge of the association between short chain fatty acids during diarrhea and changes in the microbiota composition associated with the presence of DEC pathogens.
The effect of multiple sclerosis therapy on gut microbiota dysbiosis: a longitudinal prospective study
Andreea-Cristina Paraschiv1,a, Vitalie Vacaras1,2,a, Cristina Nistor1,2, Cristiana Vacaras3, Stefan Strilciuc1 and Dafin F Muresanu1,2
The gut microbiota, a complex ecosystem with various immune functions, plays a significant role in MS, and its response to different treatments is highlighted in this study. In clinical practice, maintaining a healthy microbiota is crucial for individuals with MS.
Comparison of microbial communities and the profile of sulfate-reducing bacteria in patients with ulcerative colitis and their association with bowel diseases: a pilot study
Ivan Kushkevych1, Kristýna Martínková1, Lenka Mráková1, Francesco Giudici2, Simone Baldi2, David Novak3, Márió Gajdács4, Monika Vítězová1, Dani Dordevic5, Amedeo Amedei2 and Simon K.-M. R. Rittmann6
Considerable evidence has accumulated regarding the molecular relationship between gut microbiota (GM) composition and the onset (clinical presentation and prognosis) of ulcerative colitis UC. Our findings highlight, among other observations, significant variations in the gut microbial composition among patients with varying disease severity and activity.
Replicative aging in yeast involves dynamic intron retention patterns associated with mRNA processing/export and protein ubiquitination
Jesús Gómez-Montalvo1, Alvaro de Obeso Fernández del Valle1, Luis Fernando De la Cruz Gutiérrez1, Jose Mario Gonzalez-Meljem1 and Christian Quintus Scheckhuber1
Saccharomyces cerevisiae has yielded relevant insights into some of the basic mechanisms of organismal aging. Among these are genomic instability, oxidative stress, caloric restriction and mitochondrial dysfunction. Our work uncovers a previously unexplored layer of the transcriptional program of yeast aging and, more generally, expands the knowledge on the occurrence of alternative splicing in baker´s yeast.
Quantifying yeast lipidomics by high-performance thin-layer chromatography (HPTLC) and comparison to mass spectrometry-based shotgun lipidomics
Thorsten Meyer1, Oskar Knittelfelder2, Martin Smolnig1 and Patrick Rockenfeller1
Lipidomic analysis in diverse biological settings has become a frequent tool to increase our understanding of the processes of life. Here we describe a method to analyse basic lipidomics in yeast using HPTLC and offer comparison to measurement of the same samples with MS-based shotgun lipidomics.
A simple microfluidic platform to study age-dependent protein abundance and localization changes in Saccharomyces cerevisiae
April 13, 2017
We have developed a user-friendly microfluidic system paired with a genetic approach to enrich and study ageing mother yeast cells, enabling the monitoring of protein abundance and localization changes during the crucial first half of their replicative lifespan, leading to the discovery of novel age-dependent protein behaviors.
Thiol trapping and metabolic redistribution of sulfur metabolites enable cells to overcome cysteine overload
March 27, 2017
In this study, researchers investigate the mechanisms for handling cysteine overload using Saccharomyces cerevisiae, finding that overexpressing the high affinity cysteine transporter, YCT1, enables yeast cells to rapidly accumulate high levels of intracellular cysteine. The study demonstrates that cells can manage potentially toxic levels of cysteine by converting it to non-reactive thiol forms and utilizing the metabolic products for cell growth.
The frequency of yeast [PSI+] prion formation is increased during chronological ageing
March 27, 2017
Aging is marked by a decline in cellular functions and the increased formation of the yeast [PSI+] prion, an altered translation termination factor, which suggests that autophagy suppresses age-related prion development. Interestingly, yeast cells that adopt the [PSI+] form exhibit better survival through aging, indicating that [PSI+] formation, linked to enhanced autophagy, may confer advantages such as reduced protein aggregation and improved cell viability.
A multigene family encoding surface glycoproteins in Trypanosoma congolense
March 2, 2017
Trypanosoma congolense, the causative agent of the most important livestock disease in Africa, expresses specific surface proteins involved in its parasitic lifestyle. By mining the T. congolense genome database, we identified a novel family of lectin-like glycoproteins (TcoClecs).
Identification of Ftr1 and Zrt1 as iron and zinc micronutrient transceptors for activation of the PKA pathway in Saccharomyces cerevisiae
March 2, 2017
We now show that the yeast high-affinity iron transporter Ftr1 and high-affinity zinc transporter Zrt1 function as transceptors for the micronutrients iron and zinc. We show that replenishment of iron to iron-starved cells or zinc to zinc-starved cells triggers within 1-2 minutes a rapid surge in trehalase activity, a well-established PKA target.
Balanced CoQ6 biosynthesis is required for lifespan and mitophagy in yeast
February 3, 2017
In brief, we show that, in yeast, Ptc7 modulates the adaptation to respiratory metabolism by dephosphorylating Coq7 to supply newly synthesized CoQ6, and by activating mitophagy to remove defective mitochondria at stationary phase, guaranteeing a proper CLS in yeast.
Mutational analysis of fructose-1,6-bis-phosphatase FBP1 indicates partially independent functions in gluconeogenesis and sensitivity to genotoxic stress
February 1, 2017
Our results support predicted vital roles of several fructose-1,6-bisphosphatase residues for enzymatic activity and led to the identification of residues indispensable for the MMS-sensitizing effect. Despite an overlap between these two properties, careful analysis revealed two mutations, Asn75 and His324, which decouple the enzymatic activity and the MMS-sensitizing effect, indicating two distinctive biological activities linked in this key gluconeogenesis enzyme.
The copper transport-associated protein Ctr4 can form prion-like epigenetic determinants in Schizosaccharomyces pombe
January 2, 2017
Ctr4 exhibits multiple features diagnostic of other fungal prions and is the first example of a prion in fission yeast. These findings suggest that transmissible protein-based determinants of traits may be more widespread among fungi.
Improvement of biochemical methods of polyP quantification
December 29, 2016
As the main output of this evaluation we propose a straightforward and robust procedure that can be used as gold standard protocol for cellular polyP purification and determination from unicellular organisms, thus providing consistency to measurements and facilitating inter-laboratory comparisons and biological interpretation of the results.
Bax mitochondrial relocation is linked to its phosphorylation and its interaction with Bcl-xL
December 5, 2016
The heterologous expression of Bax, and other Bcl-2 family members, in the yeast Saccharomyces cerevisiae, has proved to be a valuable reporter system to investigate the molecular mechanisms underlying their interaction with mitochondria. Our data provide the molecular basis for a model of dynamic equilibrium for Bax localization and activation, regulated both by phosphorylation and Bcl-xL.