Superresolution imaging of ribosomes and RNA polymerase in live Escherichia coli cells
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Summary: Quantitative spatial distributions of ribosomes (S2‐YFP) and RNA polymerase (RNAP; β′‐yGFP) in live Escherichia coli are measured by superresolution fluorescence microscopy. In moderate growth conditions, nucleoid–ribosome segregation is strong, and RNAP localizes to the nucleoid lobes. The mean copy numbers per cell are 4600 RNAPs and 55 000 ribosomes. Only 10–15% of the ribosomes lie within the densest part of the nucleoid lobes, and at most 4% of the RNAPs lie in the two ribosome‐rich endcaps. The predominant observed diffusion coefficient of ribosomes is D ribo = 0.04 µm 2 s −1 , attributed to free mRNA being translated by one or more 70S ribosomes. We find no clear evidence of subdiffusion, as would arise from tethering of ribosomes to the DNA. The degree of DNA–ribosome segregation strongly suggests that in E. coli most translation occurs on free mRNA transcripts that have diffused into the ribosome‐rich regions. Both RNAP and ribosome radial distributions extend to the cytoplasmic membrane, consistent with the transertion hypothesis. However, few if any RNAP copies lie near the membrane of the endcaps. This suggests that if transertion occurs, it exerts a direct radially expanding force on the nucleoid, but not a direct axially expanding force.
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With ∼3 × 10^4 ribosomes per fL of cell volume and up to ∼400 K+ site-specifically bound per ribosome, ribosomes may bind up to ∼1.2 × 10^7 K+ ions (∼10% of the cell quota) in these sites alone.
“With ∼3 × 104 ribosomes per fL (10–15 liters) of cell volume,56 and up to ∼400 K+ site-specifically bound per ribosome,54 ribosomes may bind up to ∼1.2 × 107 K+ ions (∼10% of the cell quota) in these sites alone.”
From Metals in Motion: Understanding Labile Metal Pools in Bacteria (2025) · cites this paper as [56]
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Cites
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“We can then scale the total, pre-bleach YFP intensity to that of a single YFP molecule to estimate the copy numbers of ribosomes and RNAP in each cell (Taniguchi et al., 2010).”
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- Authors
- Bakshi, S., Siryaporn, A., Goulian, M., & Weisshaar, J. C.
- Affiliations
- University of Wisconsin–Madison (US)
- University of Pennsylvania (US)
- Corresponding author
- Weisshaar, James C.
- Venue
- Molecular Microbiology
- Publisher
- Wiley
- Published
- 2012-05-24
- Volume
- 85
- Issue
- 1
- Pages
- 21-38
- ISSN
- 0950-382X (ISSN-L)
- 1365-2958
- PMC
- PMC3383343
- OpenAlex
- W1883779479
- Cited by
- 456
- References
- 5
- Citations by year
- 2026: 16 · 2025: 27 · 2024: 43 · 2023: 30 · 2022: 19 · 2021: 28 · 2020: 37 · 2019: 51 · 2018: 41 · 2017: 34 · 2016: 29 · 2015: 23 · 2014: 35 · 2013: 34 · 2012: 9
- FWCI
- 12.15
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Molecular Microbiology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Cambridge University Engineering Department Publications Database (submittedVersion)
- Funders
- National Science Foundation (0615957)
- National Institute of General Medical Sciences (R01GM086468, R01GM080279, R01GM094510)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Keywords
- Escherichia coli, plasma membrane
- MeSH terms
- DNA, Bacterial, Escherichia coli, Image Processing, Computer-Assisted, Microscopy, Fluorescence, Ribosomes, DNA-Directed RNA Polymerases, Escherichia coli Proteins
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