A5091397008- Fungal and yeast genetics research
- Mitochondrial Function and Pathology
- CRISPR and Genetic Engineering
- RNA and protein synthesis mechanisms
- Microbial Metabolic Engineering and Bioproduction
- Genomics and Chromatin Dynamics
- Ubiquitin and proteasome pathways
- Genomics and Phylogenetic Studies
- Photosynthetic Processes and Mechanisms
- Advanced Proteomics Techniques and Applications
- Biofuel production and bioconversion
- Cell Image Analysis Techniques
- Evolutionary Game Theory and Cooperation
- Bioinformatics and Genomic Networks
- Evolution and Genetic Dynamics
- Protein Degradation and Inhibitors
- DNA Repair Mechanisms
- Advanced Fluorescence Microscopy Techniques
- Enzyme Structure and Function
- Lipid metabolism and biosynthesis
- Genetically Modified Organisms Research
Weizmann Institute of Science
2017-2025
University of Copenhagen
2014
Hebrew University of Jerusalem
1998-2006
A single translation product of the<i>FUM</i>1 gene encoding fumarase is distributed between the cytosol and mitochondria <i>Saccharomyces cerevisiae</i>. All products are targeted processed in before distribution. Here we show that targeting coupled to initially involves insertion protein across mitochondrial membranes processing by matrix protease. Rapid folding may determine its requirement for coupling translocation with unique route The amino termini most molecules translocated...
The ability to measure the abundance and visualize localization of proteins across yeast proteome has stimulated hypotheses on gene function fueled discoveries. While classic C' tagged GFP library been only resource for over a decade, recent development SWAT technology led creation multiple novel libraries where new-generation fluorescent reporters are fused at N' open reading frames. Efficient access these data requires user interface compare protein abundance, co-localization cells,...
We have previously proposed that a single translation product of the FUM1 gene encoding fumarase is distributed between cytosol and mitochondria Saccharomyces cerevisiae all products are targeted processed in before distribution. Alternative models for distribution been require more than one product. In current work (i) we show by using sequential Edman degradation mass spectrometry cytosolic mitochondrial isoenzymes an identical amino terminus formed cleavage processing peptidase, (ii)...
We have previously proposed that a single translation product of the FUM1 gene encoding fumarase is distributed between cytosol and mitochondria Saccharomyces cerevisiae all products are targeted processed in before distribution. Thus, returns to cytosol. In current work, we (i) generated mutations throughout coding sequence which resulted fumarases with altered conformations but lost their ability be distributed; (ii) showed by mass spectrometry mature cytosolic mitochondrial isoenzymes...
In experimental evolution, scientists evolve organisms in the lab, typically by challenging them to new environmental conditions. How best a desired trait? Should challenge be applied abruptly, gradually, periodically, sporadically? one apply chemical mutagenesis, and do strains with high innate mutation rate faster? What are ideal population sizes of evolving populations? There endless strategies, beyond those that can exposed individual labs. We therefore arranged community challenge,...
A significant challenge in cell biology is to uncover the function of uncharacterized proteins. Surprisingly a quarter proteome still poorly understood even most well studied model organisms. Systematic methodologies, including use tagged protein collections, have emerged as powerful approach address this gap. Despite availability proteome-wide collections featuring various fused proteins, impact tag size on highlighted need for using minimally disruptive tags functional genomic studies. To...
Abstract Genome-wide collections of yeast strains, known as libraries, revolutionized the way systematic studies are carried out. Specifically, libraries that involve a cellular perturbation, such deletion collection, have facilitated key biological discoveries. However, short-term rewiring and long-term accumulation suppressor mutations often obscure functional consequences perturbations. We present AID library which supplies “on demand” protein depletion to overcome these limitations....
Abstract Clone collections of modified strains (“libraries”) are a major resource for systematic studies with the yeast Saccharomyces cerevisiae . Construction such libraries is time-consuming, costly and confined to genetic background specific strain. To overcome these limitations, we present CRISPR-Cas12a (Cpf1)-assisted tag library engineering (CASTLING) multiplexed strain construction. CASTLING uses microarray-synthesized oligonucleotide pools in vitro recombineering program genomic...
Genome-wide collections of yeast strains, known as libraries, revolutionized the way systematic studies are carried out. Specifically, libraries that involve a cellular perturbation, such deletion collection, have facilitated key biological discoveries. However, short-term rewiring and long-term accumulation suppressor mutations often obscure functional consequences perturbations. We present AID library which supplies “on demand” protein depletion to overcome these limitations. Here, each is...
Here we describe a C-SWAT library for high-throughput tagging of Saccharomyces cerevisiae ORFs. It consists 5661 strains with an acceptor module inserted after each ORF, which can be efficiently replaced tags or regulatory elements. We validate the targeted sequencing and demonstrate its use by yeast proteome bright fluorescent proteins, determining how sequences downstream ORFs influence protein expression localizing previously undetected proteins.
Abstract Clone collections of modified strains (‘libraries’) are a major resource for systematic studies with the yeast Saccharomyces cerevisiae. Construction such libraries is time-consuming, costly and confined to genetic background specific strain. To overcome these limitations, we present CRISPR-Cas12a (Cpf1)-assisted tag library engineering (CASTLING) multiplexed strain construction. CASTLING uses microarray-synthesized oligonucleotide pools in vitro recombineering program genomic...