Genetic Mapping of MAPK-Mediated Complex Traits Across S. cerevisiae

Abstract

Signaling pathways enable cells to sense and respond to their environment. Many cellular signaling strategies are conserved from fungi to humans, yet their activity and phenotypic consequences can vary extensively among individuals within a species. A systematic assessment of the impact of naturally occurring genetic variation on signaling pathways remains to be conducted. In S. cerevisiae, both response and resistance to stressors that activate signaling pathways differ between diverse isolates. Here, we present a quantitative trait locus (QTL) mapping approach that enables us to identify genetic variants underlying such phenotypic differences across the genetic and phenotypic diversity of S. cerevisiae. Using a Round-robin cross between twelve diverse strains, we identified QTL that influence phenotypes critically dependent on MAPK signaling cascades. Genetic variants under these QTL fall within MAPK signaling networks themselves as well as other interconnected signaling pathways. Finally, we demonstrate how the mapping results from multiple strain background can be leveraged to narrow the search space of causal genetic variants.

The Hidden Complexity of Mendelian Traits across Natural Yeast Populations

“…Yeasts and more particularly S. cerevisiae have been extensively used as a model for dissecting many complex traits that were of medical, industrial and evolutionary interests (Bloom et al, 2013; Ehrenreich et al, 2012; Mukherjee et al, 2014; Steinmetz et al, 2002; Treusch et al, 2015). A trend emerging from studying complex traits in this species was that causal variants do not distribute randomly across the genome, and several hotspots have been identified (Fay, 2013).…”

“…Fragment containing PDR1 and its native promoter and terminator regions flanked by attB1/attB2 recombination sites was amplified from the genomic DNA of YJM326 and Σ1278b, and was cloned into an empty centromeric plasmid with HphMX resistance marker (pCTRL, Treusch et al, 2015), according to instructions. The resulting plasmids, pPDR1 YJM326 and pPDR1 Σ1278b were verified using restriction enzymes and PCR amplification with internal primers of PDR1 gene.…”

The high osmotic response and cell wall integrity pathways cooperate to regulate morphology, microsclerotia development, and virulence in Metarhizium rileyi

“…Fungi contain complex signalling pathway networks to handle with these stresses15. Mitogen-activated protein kinase (MAPK) signalling pathways are conserved and play important roles in sensing environmental stimuli, in transmitting extracellular signals to the nucleus to modulate gene expression, in regulating morphology, in responding to abiotic and biotic stresses, and in virulence/pathogenicity161718.…”

Molecular Identification of d-Ribulokinase in Budding Yeast and Mammals

“…The Gateway plasmid pDEST527 was a gift from Dominic Esposito (Addgene plasmid 11518). The empty Gateway plasmid p41Hyg 1-F GW was a gift from Leonid Kruglyak and Sebastian Treusch (Addgene plasmid 58547) (63).…”

“…A plasmid (p41Hyg 1-F GW-YDR109C) to rescue the yeast metabolic phenotype after YDR109C deletion was prepared by Gateway insertion of the YDR109C coding sequence as well as the 840 nucleotides upstream of the ATG start codon and 300 nucleotides downstream of the stop codon into the low copy number CEN vector p41Hyg 1-F GW (63). The YDR109C gene sequence was PCR-amplified from prototrophic wild-type S. cerevisiae FY4 (MATa) genomic DNA using the YDR109cFwdres and YDR109cRevres primers (supplemental Table S5).…”