Skip to main content
. Author manuscript; available in PMC: 2021 Mar 15.
Published in final edited form as: Mol Cell. 2020 Nov 19;81(1):139–152.e10. doi: 10.1016/j.molcel.2020.10.035

Figure 5. Test of importance of UP elements in activity of PigR-regulated promoters.

Figure 5.

A Mutations were made in the PRE (PRE mutant 1 and PRE mutant 3 in bold) as well as in the UP element downstream of the PRE (UP mutant 1, in bold). Wild type is also labeled. In PRE mutant 3, the TA bps of the PRE at positions 6 and 7 are mutated to CG. In UP mutant 1, bps 2 and 3 in the UP element downstream of the PRE were changed from AA to GC. All mutations impaired PigR-dependent regulation. B Quantification of iglA-lacZ expression in LVS wild-type (LVS) and ΔpigR mutant (LVS ΔpigR) cells containing the indicated iglA promoter variants (X-axis) by β-galactosidase assay (Miller units). Promoter variants linked to a lacZ reporter gene were integrated into the FTL_0111 locus. Statistical significance was assessed in Prism using one-way ANOVA with Tukey’s multiple comparisons; **** P<0.0001. C Double mutations were made in the PRE (PRE mutant 3, as indicated in bold) and in the UP element downstream of the PRE (UP mutant 1, as indicated in bold). Wild type is labeled. In PRE mutant 3, the TA bps of the PRE at positions 6 and 7 are mutated to CG. In UP mutant 1, bps 2 and 3 in the FTL_0026 promoter downstream UP element were changed from AA to GC. All mutations impaired PigR-dependent regulation. D Quantification of FTL_0026-lacZ expression in LVS wild-type (LVS) and ΔpigR mutant (LVS ΔpigR) cells containing the indicated FTL_0026 promoter variants (X-axis) by β-galactosidase assay (Miller units). Promoter variants linked to a lacZ reporter gene were integrated into the FTL_0026 locus. Statistical significance was assessed in Prism using one-way ANOVA with Tukey’s multiple comparisons; **** P<0.0001.