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GC-Rich DNA Elements Enable Replication Origin Activity in the Methylotrophic Yeast Pichia pastoris

Figure 3

Deep mutational scanning of P. pastoris ARSs.

(A) Schematic of the mutARS-seq deep mutational scanning experiment. Auxotrophic ura3 yeast were transformed with a library of mutant ARS variants and competed in selective medium. The abundance of different ARS variants was determined by deep sequencing at intervals during competitive growth. (B) Results of mutARS-seq of ARS-C379. The relevant sequence of ARS-C379 is shown with the best match to the GC-ACS motif highlighted in red (and a 3′ constrained dinucleotide highlighted in blue). The log-transformed enrichment ratio is shown for each nucleotide at each position along the sequence. (C) Results of mutARS-seq of ARS-A2772. Same as in (B), except that the motif logo shown was constructed from the enrichment ratio scores post-analysis, whereas the motif shown in (B) was constructed from ARS alignments.

Figure 3