The effect of autonomously replicating sequences on gene expression in saccharomyces cerevisiae

Autonomously replicating sequences (ARSs) are essential for genome replication and can regulate gene expression as potential cis-elements; however, the mechanisms underlying this regulation are unclear. In order to understand these mechanisms, we first replaced genes adjacent to ARSs on chromosome III of Saccharomyces cerevisiae with an expression cassette containing the gene encoding red fluorescence protein (RFP) and verified that the relative fluorescence intensity of RFP was positively correlated with the activities of the adjacent ARSs. Then, to investigate the effect of ARSs on gene expression, we designed ARS-insertion and -deletion variants, finding that the insertion of a strongly active ARS adjacent to a weakly active ARS enhanced adjacent gene expression, whereas the deletion of a strongly active ARS reduced the expression of the flanking RFP; however, insertion of multiple strong ARSs did not show a synergistic effect on gene expression. Further analysis suggested that ARSs located in compact chromosomal regions exerted a stronger effect on gene expression than those in relaxed chromosomal regions. These results demonstrated that ARSs regulated S. cerevisiae gene expression by interacting with the chromosome structure and provide a potential strategy for regulating gene expression in synthetic biology and metabolic engineering studies using S. cerevisiae.