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. Author manuscript; available in PMC: 2009 Oct 27.
Published in final edited form as: Mol Microbiol. 2009 May 25;73(4):519–533. doi: 10.1111/j.1365-2958.2009.06744.x

Fig. 5.

Fig. 5

The activities of the V132A and L138A variants are rescued by ADP–AlF.

A. SDS-PAGE gel showing the cross-linking profiles of σ54–DNA complexes formed on the mismatch promoter probe in the presence of ADP–AlF. The migration positions of the cross-linked σ54–DNA and PspF1–275–DNA species are indicated. A cross-linked PspF1–275–DNA species is observed in reactions containing the V132A (lane 4), S135A (lane 6), Q136A (lane 7), P137A/T (lanes 8–9) and L138A (lane 10) variants. Native-PAGE gel illustrating that stable ADP–AlF trapped complexes are only observed in the presence of PspF1–275 WT (lane 2) and the S135A (lane 6), Q136A (lane 7) and P137A/T (lanes 8–9) variants. The migration positions of the σ54–DNA-PspF1–275:ADP–AlF (trapped) and binary σ54–DNA (σ54–DNA) complexes, free DNA and percentage DNA bound in each complex are as indicated.

B. SDS-PAGE gel as in A but on the duplex promoter probe in the presence of core RNAP. The migration positions of the cross-linked β/β′–DNA, σ54–DNA and PspF1–275–DNA species are indicated.