Figure 7.

GroEL inhibits σ³²-dependent in vitro transcription. Multiround in vitro transcription was performed with holoenzyme containing either σ³² (Eσ³²) or σ⁷⁰ (Eσ⁷⁰) (100 nM) incubated with GroEL (500 nM), GroES (1 μM), GroELY203E (500 nM), DnaK (2 μM), DnaJ (400 nM), or combinations thereof. An end-labeled oligo was added to each reaction as an internal control. Transcription reactions were phenol-chloroform extracted, ethanol precipitated, and analyzed on a 6% polyacrylamide gel. (A) Representative transcription gel showing duplicate reactions documenting GroEL inhibition of Eσ³². (B) Quantification and summary of transcription results from A as well as from additional transcription experiments.