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NusA and NusG are transcription elongation factors that bind to RNA polymerase (RNAP) after σ subunit release.Escherichia coli NusA (NusAEc ) stimulates intrinsic termination and RNAPEc pausing, whereas NusGEc promotes Rho-dependent termination and pause escape. Both Nus factors also participate in the formation of RNAPEc antitermination complexes. We showed thatBacillus subtilis NusA (NusABs ) stimulates intrinsic termination and RNAPBs pausing at U107 and U144 in thetrpEDCFBA operon leader. Pausing at U107 and U144 participates in the transcription attenuation and translational control mechanisms, respectively, presumably by providing additional time fortrp RNA-binding attenuation protein (TRAP) to bind to the nascenttrp leader transcript. Here, we show that NusGBs causes modest pause stimulation at U107 and dramatic pause stimulation at U144. NusABs and NusGBs act synergistically to increase the U107 and U144 pause half-lives. NusGBs -stimulated pausing at U144 requires RNAPBs , whereas NusABs stimulates pausing of RNAPBs and RNAPEc at the U144 andE. coli his pause sites. Although NusGEc does not stimulate pausing at U144, it competes with NusGBs -stimulated pausing, suggesting that both proteins bind to the same surface of RNAPBs . Inactivation ofnusG results in the loss of RNAP pausing at U144in vivo and elevatedtrp operon expression, whereas plasmid-encoded NusG complements the mutant defects. Overexpression ofnusG reducestrp operon expression to a larger extent than overexpression ofnusA .

Function of the Bacillus subtilis transcription elongation factor NusG in hairpin-dependent RNA polymerase pausing in the trp leader