Fig. 8: Identification of tlh genes with multiple mutations.

a Identification of tlh3 and tlh4. Chromosomal locations of the four tlh genes are shown. b Summary of predicted ORFs and nucleotide changes in the tlh genes of 972SD4 compared with the tlh2 gene in PomBase. The top panel shows the tlh2 locus in PomBase. Seven digit numbers indicate chromosomal positions on Ch2 in PomBase. Blue arrow with a purple line, the tlh2 ORF defined in PomBase; brown bar, dh-homologous region; light purple line, predicted coding region which is in-frame with that of the tlh2 ORF in PomBase; light or dark green line, predicted coding region in the reading frame different from that of the predicted tlh2 ORF in PomBase; yellow star, point mutation (no frame shift); orange arrowhead, nucleotide insertion (numbers of inserted nucleotides are shown below); blue arrowhead, nucleotide deletion (numbers of deleted nucleotides are shown below); dM the original first methionine codon of the tlh2 ORF in PomBase, uM putative first methionine codon upstream of dM for PomBase-tlh2, uM′ putative first methionine codon for the short ORF, dM′ putative first methionine codon for the long ORF, T termination codon. c Expression of RNA of the dh-homologous region in the tlh genes in strains 972, SD5, and 972SD4 in a clr4∆ background was analyzed by quantitative RT-PCR in the presence (+) or absence (−) of RNase. Expression level of tlh⁺ relative to that of his1⁺ was normalized to that in 972 (−RNase). Data are presented as mean values ± SD (standard deviations). N = 3 biologically independent experiments (each value is indicated by a gray dot).