Fig. 6: The CcpA HTH domain is required for efficient RNA-binding in vivo. | Nature Communications

Fig. 6: The CcpA HTH domain is required for efficient RNA-binding in vivo.

From: The RNA-bound proteome of MRSA reveals post-transcriptional roles for helix-turn-helix DNA-binding and Rossmann-fold proteins

Fig. 6

a Crystal structure of a CcpA-HPr-dsDNA complex (grey, yellow and blue, respectively) containing a 16 bp CRE dsDNA substrate (adapted from34; PDB code 1RZR https://www.wwpdb.org/pdb?id=pdb_00001rzr). The zoomed-in region shows the location of two threonines that are required for dsDNA binding. HPr histidine-containing protein, CRE Catabolite Response Elements. b The CcpA T18DT33D mutation causes a growth defect. Growth curves were performed in a plate reader using the following strains: USA300 + pCN33-PtufA, USA300 ΔccpA, USA300 ΔccpA + pCN33-PtufA::ccpA and the USA300 ΔccpA + pCN33-PtufA::ccpAT18DT33D. Data are presented as mean values (solid lines) +/− SD (shaded region). c T18 and T33 are required for efficient RNA-binding in vivo. Shown is a result from a CcpA CRAC experiment using the ΔccpA as a negative control, the WT CcpA-HTF protein as positive control and the CcpA T18DT33D-HTF mutant. d Quantification of CcpA-HTF WT and T18DT33D cross-linking data. Autoradiography and Western blot signals from two independently performed CRAC experiments (shown in c) were quantified, and the autoradiography signal was normalised to the CcpA protein levels. Original data and pictures for b–d are provided in Source data.

Back to article page