Multiscale spatial organization of RNA polymerase in Escherichia coli.
Nucleic acid synthesis is spatially organized in many organisms. In bacteria, however, the spatial distribution of transcription remains obscure, owing largely to the diffraction limit of conventional light microscopy (200-300 nm). Here, we use photoactivated localization microscopy to localize indi...
| Main Authors: | , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2013
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| _version_ | 1826288989975347200 |
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| author | Endesfelder, U Finan, K Holden, S Cook, P Kapanidis, A Heilemann, M |
| author_facet | Endesfelder, U Finan, K Holden, S Cook, P Kapanidis, A Heilemann, M |
| author_sort | Endesfelder, U |
| collection | OXFORD |
| description | Nucleic acid synthesis is spatially organized in many organisms. In bacteria, however, the spatial distribution of transcription remains obscure, owing largely to the diffraction limit of conventional light microscopy (200-300 nm). Here, we use photoactivated localization microscopy to localize individual molecules of RNA polymerase (RNAP) in Escherichia coli with a spatial resolution of ∼40 nm. In cells growing rapidly in nutrient-rich media, we find that RNAP is organized in 2-8 bands. The band number scaled directly with cell size (and so with the chromosome number), and bands often contained clusters of >70 tightly packed RNAPs (possibly engaged on one long ribosomal RNA operon of 6000 bp) and clusters of such clusters (perhaps reflecting a structure like the eukaryotic nucleolus where many different ribosomal RNA operons are transcribed). In nutrient-poor media, RNAPs were located in only 1-2 bands; within these bands, a disproportionate number of RNAPs were found in clusters containing ∼20-50 RNAPs. Apart from their importance for bacterial transcription, our studies pave the way for molecular-level analysis of several cellular processes at the nanometer scale. |
| first_indexed | 2024-03-07T02:22:04Z |
| format | Journal article |
| id | oxford-uuid:a448c232-5691-42e4-87dd-5b8662d381f7 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T02:22:04Z |
| publishDate | 2013 |
| record_format | dspace |
| spelling | oxford-uuid:a448c232-5691-42e4-87dd-5b8662d381f72022-03-27T02:32:49ZMultiscale spatial organization of RNA polymerase in Escherichia coli.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:a448c232-5691-42e4-87dd-5b8662d381f7EnglishSymplectic Elements at Oxford2013Endesfelder, UFinan, KHolden, SCook, PKapanidis, AHeilemann, MNucleic acid synthesis is spatially organized in many organisms. In bacteria, however, the spatial distribution of transcription remains obscure, owing largely to the diffraction limit of conventional light microscopy (200-300 nm). Here, we use photoactivated localization microscopy to localize individual molecules of RNA polymerase (RNAP) in Escherichia coli with a spatial resolution of ∼40 nm. In cells growing rapidly in nutrient-rich media, we find that RNAP is organized in 2-8 bands. The band number scaled directly with cell size (and so with the chromosome number), and bands often contained clusters of >70 tightly packed RNAPs (possibly engaged on one long ribosomal RNA operon of 6000 bp) and clusters of such clusters (perhaps reflecting a structure like the eukaryotic nucleolus where many different ribosomal RNA operons are transcribed). In nutrient-poor media, RNAPs were located in only 1-2 bands; within these bands, a disproportionate number of RNAPs were found in clusters containing ∼20-50 RNAPs. Apart from their importance for bacterial transcription, our studies pave the way for molecular-level analysis of several cellular processes at the nanometer scale. |
| spellingShingle | Endesfelder, U Finan, K Holden, S Cook, P Kapanidis, A Heilemann, M Multiscale spatial organization of RNA polymerase in Escherichia coli. |
| title | Multiscale spatial organization of RNA polymerase in Escherichia coli. |
| title_full | Multiscale spatial organization of RNA polymerase in Escherichia coli. |
| title_fullStr | Multiscale spatial organization of RNA polymerase in Escherichia coli. |
| title_full_unstemmed | Multiscale spatial organization of RNA polymerase in Escherichia coli. |
| title_short | Multiscale spatial organization of RNA polymerase in Escherichia coli. |
| title_sort | multiscale spatial organization of rna polymerase in escherichia coli |
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