A new laboratory evolution approach to select for constitutive acetic acid tolerance in Saccharomyces cerevisiae and identification of causal mutations

Table 3 Mutations present in multiple evolved S. cerevisiae strains and genetic analysis of their relevance for acetic acid tolerance

Gene	Strain	SNP	Nucleotide in THS	Nucleotide in SHS
ASG1	MUT1A	G1248A	A	G
	MUT2B	G1248T	T	G
	HAT1E	A1979G	G	A
	HAT2A	G2881C	C	G
ADH3	MUT1A	G416T	T	G
	MUT2B	T966G	T-G	T
	HAT2A	T201A	A	T
SKS1	MUT2B	G821T	T	G
SKS1	HAT2A	C617A	C-A	nd
GIS4	MUT3E	G1322C	C	G
GIS4	UV-E3	G295A	nd	nd
SAC6	MUT1A	G491A	A	G
	MUT2B	G491A	nd	A
	MUT3E	G491A	G-A	G
	HAT1E	G491A	G	A
	HAT2A	G491A	G-A	A
EUG1	MUT1A	G-113T	G	G-T
	MUT2B	G-113T	G-T	T
	MUT3E	G-113T	G	T
	HAT1E	G-113T	G-T	G
	HAT2A	G-113T	T	T

Mutations were identified by comparing whole-genome sequencing data of single-cell isolates MUT1A, MUT2B, MUT3E, HAT1E, HAT2A and UV-E3 to the genome sequence of S. cerevisiae CEN.PK113-7D [44]. Mutations shown occur in at least two strains. In addition to the nucleotide changes, the distribution of the SNP among tolerant haploid segregants (THS) and the sensitive haploid segregants (SHS) of a diploid, generated by mating the tolerant strains with S. cerevisiae IMK439, are shown. Nucleotides present in the evolved strains are bold and underlined. Two nucleotides separated by a dash indicate presence of both mutated and reference-strain alleles in pooled THS or SHS. nd no sequence data obtained

ISSN: 2731-3654