Extended Data Fig. 3: EMSA data for release of wild-type and mutant MmfR from the mmyB–mmyY and mmfL–mmfR intergenic regions in the presence of increasing quantities of the MMFs, MMF analogues and SCB1.
From: Molecular basis for control of antibiotic production by a bacterial hormone
a, Interaction of MmfR with DNA fragments corresponding to the mmyB–mmyY intergenic region (230 bp) in response to increasing amounts of MMFs. b, Interaction of MmfR with DNA fragments corresponding to the mmfL–mmfR intergenic region (194 bp) in response to increasing amounts of MMFs. c, Interaction of MmfR with the DNA fragments corresponding to the mmfL–mmfR intergenic region (194 bp) in response to increasing amounts of SCB1. d, EMSAs showing that Y85 and Q130 of MmfR have an important role in hormone-induced DNA release. Approximately ten times the quantity of MMF1 is required to release the Y85F and Q130E mutants of MmfR from the mmfL–mmfR intergenic region (194 bp) than the wild-type protein. e, Interaction of MmfR with the DNA fragments corresponding to the mmfL–mmfR intergenic region (194 bp) in response to increasing amounts of synthetic MMF analogues. Lane 1, isolated DNA fragments (0.1 pmol); lane 2, DNA fragments (0.1 pmol) mixed with protein (1.8 pmol). For a, b and d, lanes 3 to 9 show addition of increasing quantities of MMFs (0.8, 4, 8, 14, 20, 40 and 100 nmol respectively) to the protein–DNA complexes. For c, lanes 3 to 7 show addition of increasing quantities of racemic SCB1 (0.8, 8, 20, 40 and 400 nmol respectively) to the DNA–protein complexes. For e, lanes 3 to 6 show addition of increasing quantities of MMF analogues (0.8, 8, 20 and 100 nmol respectively) to the DNA–protein complexes. At least two independent technical replicates of each EMSA were conducted and in all cases similar results were obtained. For gel source data, see Supplementary Fig. 1a–d.
