Computational and experimental assessment of key interdomain residues controlling the fold-switch of RfaH.

ABSTRACT: Escherichia coli RfaH, a member of the universally conserved NusG family of transcription factors, regulates its function by undergoing a structural rearrangement of its C-terminal domain (CTD) upon recruitment to RNA polymerase (RNAP) paused at the DNA signal known as operon polarity suppressor (ops) element. While it is known that the fold-switch of RfaH CTD from an α-helical hairpin (autoinhibited state) into a β-barrel (active state) is controlled by interactions between the N-terminal domain (NTD) and CTD, which are broken apart upon NTD binding to RNAP, a comprehensive analysis of residues that stabilize the autoinhibited state is lacking. Here, we utilize a combination of molecular dynamics (MD), protein structure prediction, and in vivo functional assays as a workflow to determine

SUBMITTER: Tabilo-Agurto C 

PROVIDER: S-EPMC12168481 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature