Comprehensive Synthetic Genetic Array Analysis of Alleles That Interact with Mutation of the Saccharomyces cerevisiae RecQ Helicases Hrq1 and Sgs1
| Autor: | Phoebe A Nguyen, Matthew L. Bochman, Cody M. Rogers, Elsbeth Sanders |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
DNA repair
RecQ helicase Saccharomyces cerevisiae Biology yeast QH426-470 03 medical and health sciences chemistry.chemical_compound Genetics saccharomyces cerevisiae Molecular Biology Genetics (clinical) 030304 developmental biology 0303 health sciences 030302 biochemistry & molecular biology DNA replication Helicase Synthetic genetic array biology.organism_classification hrq1 dna helicase chemistry biology.protein sgs1 DNA Sgs1 |
| Zdroj: | G3: Genes, Genomes, Genetics, Vol 10, Iss 12, Pp 4359-4368 (2020) |
| ISSN: | 2160-1836 |
| DOI: | 10.1534/g3.120.401709 |
| Popis: | Most eukaryotic genomes encode multiple RecQ family helicases, including five such enzymes in humans. For many years, the yeast Saccharomyces cerevisiae was considered unusual in that it only contained a single RecQ helicase, named Sgs1. However, it has recently been discovered that a second RecQ helicase, called Hrq1, resides in yeast. Both Hrq1 and Sgs1 are involved in genome integrity, functioning in processes such as DNA inter-strand crosslink repair, double-strand break repair, and telomere maintenance. However, it is unknown if these enzymes interact at a genetic, physical, or functional level as demonstrated for their human homologs. Thus, we performed synthetic genetic array (SGA) analyses of hrq1Δ and sgs1Δ mutants. As inactive alleles of helicases can demonstrate dominant phenotypes, we also performed SGA analyses on the hrq1-K318A and sgs1-K706A ATPase/helicase-null mutants, as well as all combinations of deletion and inactive double mutants. We crossed these eight query strains (hrq1Δ, sgs1Δ, hrq1-K318A, sgs1-K706A, hrq1Δ sgs1Δ, hrq1Δ sgs1-K706A, hrq1-K318A sgs1Δ, and hrq1-K318A sgs1-K706A) to the S. cerevisiae single gene deletion and temperature-sensitive allele collections to generate double and triple mutants and scored them for synthetic positive and negative genetic effects based on colony growth. These screens identified hundreds of synthetic interactions, supporting the known roles of Hrq1 and Sgs1 in DNA repair, as well as suggesting novel connections to rRNA processing, mitochondrial DNA maintenance, transcription, and lagging strand synthesis during DNA replication. |
| Databáze: | OpenAIRE |
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