Abstract
RNA polymerase (RNAP) has been shown to exhibit clustered distribution in bacterial cells. These clusters were hypothesized to be active transcription factories in which concentrated RNAP molecules are engaged in ribosomal RNA synthesis. To examine this hypothesis, in this study we used photoactivated localization microscopy (PALM) to map the spatial distribution of RNAP in correlation with active transcription in superresolution. We monitored the colocalization of RNAP with single mRNA or rRNA gene sites, with nascent transcripts, and with elongation factors NusA and NusB. We further examined the cellular positioning of rRNA and mRNA genes sharing similar regulatory controls under different transcription conditions. Our results provide evidence rigorously examining the transcription factory model.
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