Abstract
Transcription by RNA polymerase (RNAP) is coupled with translation in bacteria. Here, we observe the dynamics of transcription and subcellular localization of a specific gene locus (encoding a non-membrane protein) in living E. coli cells at subdiffraction-limit resolution. The movement of the gene locus to the nucleoid periphery correlates with transcription, driven by either E. coli RNAP or T7 RNAP, and the effect is potentiated by translation.
Highlights
Transcription by RNA polymerase (RNAP) is coupled with translation in bacteria
Using T7 RNAP transcription system that is uncoupled with translation, we demonstrate that two factors are involved in gene locus movement during transcription and responsible for the relocation of gene loci during transcription to the nucleoid periphery: transcriptional activity and ribosome binding to mRNAs
In order to visualize the location of a specific gene locus occupied by RNAPs, we replaced the endogenous promoter of the lac operon with a T7 RNAP-specific promoter (Fig. 1a)
Summary
Transcription by RNA polymerase (RNAP) is coupled with translation in bacteria. Here, we observe the dynamics of transcription and subcellular localization of a specific gene locus (encoding a non-membrane protein) in living E. coli cells at subdiffraction-limit resolution. The movement of the gene locus to the nucleoid periphery correlates with transcription, driven by either E. coli RNAP or T7 RNAP, and the effect is potentiated by translation. The complex structure of the nucleoid and the active reactions of other proteins, such as DNA polymerases, other RNAPs, and ribosomes, on the same DNA and mRNA molecules may affect transcription reactions in living cells[11]. It is well known that Escherichia coli (E. coli) RNAPs form distinct foci located at the nucleoid periphery under fast growth conditions[20,22,23] These foci have been suggested to actively transcribe genes, such as rrn operons[20,23,24].
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