Home LiteratureArticle Details
PMID: 10675340 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Compartmentalization of transcription and translation in Bacillus subtilis.

The EMBO journal ·Vol. 19 ·No. 4 ·2000-02-15 ·Pages 710-8

Lewis PJ, Thaker SD, Errington J

Abstract

Using fusions of green fluorescent protein to subunits of RNA polymerase (RNAP) and ribosomes, we have investigated the subcellular localization of the transcriptional and translational machinery in the bacterium Bacillus subtilis. Unexpectedly, we found that RNAP resides principally within the nucleoid. Conversely, ribosomes localized almost exclusively outside the nucleoid, concentrating particularly towards sites of cell division. This zonal localization was not dependent on cell division and is probably due, at least in part, to exclusion from the nucleoid. Dual labelling of RNAP and ribosomes was used to confirm the spatial separation of the two processes. We conclude that, even in the absence of a nuclear membrane, transcription and translation occur predominantly in separate functional domains. At higher growth rates, concentrations of RNAP developed, probably representing the sites of rRNA synthesis. These may represent a further spatial specialization, possibly equivalent to the eukaryotic nucleolus.

MeSH Terms
Bacillus subtilis/genetics,metabolism Base Sequence Cell Compartmentation/genetics DNA Primers/genetics DNA-Directed RNA Polymerases/genetics,metabolism Green Fluorescent Proteins Luminescent Proteins/genetics,metabolism Protein Biosynthesis RNA, Bacterial/biosynthesis RNA, Ribosomal/biosynthesis Recombinant Fusion Proteins/genetics,metabolism Ribosomes/metabolism Transcription, Genetic
Chemicals
DNA Primers Luminescent Proteins RNA, Bacterial RNA, Ribosomal Recombinant Fusion Proteins Green Fluorescent Proteins DNA-Directed RNA Polymerases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Lewis P J
Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK. [email protected]
Thaker S D
Errington J
References (36)
36 references, click to expand
  1. Cloning and characterization of a relA/spoT homologue from Bacillus subtilis.
    Mol Microbiol. 1997 Oct;26(1):65-79 PMID: 9383190
  2. REQUIREMENTS FOR TRANSFORMATION IN BACILLUS SUBTILIS.
    J Bacteriol. 1961 May;81(5):741-6 PMID: 16561900
  3. Changing views on the nature of the bacterial cell: from biochemistry to cytology.
    J Bacteriol. 1999 Jul;181(14):4143-5 PMID: 10400568
  4. Shape and fine structure of nucleoids observed on sections of ultrarapidly frozen and cryosubstituted bacteria.
    J Bacteriol. 1985 Jun;162(3):960-71 PMID: 3922958
  5. Bacterial cell division and the Z ring.
    Annu Rev Biochem. 1997;66:93-116 PMID: 9242903
  6. Altered arrangement of proteins in the spore coat of a germination mutant of Bacillus subtilis.
    J Gen Microbiol. 1983 Jun;129(6):1945-58 PMID: 6415224
  7. The Bacillus subtilis DivIVA protein targets to the division septum and controls the site specificity of cell division.
    Mol Microbiol. 1997 Jun;24(5):905-15 PMID: 9219999
  8. Dynamic movement of the ParA-like Soj protein of B. subtilis and its dual role in nucleoid organization and developmental regulation.
    Mol Cell. 1999 Nov;4(5):673-82 PMID: 10619015
  9. Dynamic, mitotic-like behavior of a bacterial protein required for accurate chromosome partitioning.
    Genes Dev. 1997 May 1;11(9):1160-8 PMID: 9159397
  10. Initiation of mRNA translation in prokaryotes.
    Biochemistry. 1990 Jun 26;29(25):5881-9 PMID: 2200518
  11. Novel roles for classical factors at the interface between translation termination and initiation.
    Mol Cell. 1999 May;3(5):601-9 PMID: 10360176
  12. FtsZ in Bacillus subtilis is required for vegetative septation and for asymmetric septation during sporulation.
    Genes Dev. 1991 Mar;5(3):447-55 PMID: 1848202
  13. Transcription mapping of the Escherichia coli chromosome by electron microscopy.
    J Bacteriol. 1989 Aug;171(8):4207-16 PMID: 2666391
  14. Localization of bacterial DNA polymerase: evidence for a factory model of replication.
    Science. 1998 Nov 20;282(5393):1516-9 PMID: 9822387
  15. Bacillus subtilis cell cycle as studied by fluorescence microscopy: constancy of cell length at initiation of DNA replication and evidence for active nucleoid partitioning.
    J Bacteriol. 1998 Feb;180(3):547-55 PMID: 9457856
  16. Bacterial cell division: a moveable feast.
    Proc Natl Acad Sci U S A. 1999 May 25;96(11):5891-3 PMID: 10339512
  17. A fixed distance for separation of newly replicated copies of oriC in Bacillus subtilis: implications for co-ordination of chromosome segregation and cell division.
    Mol Microbiol. 1998 Jun;28(5):981-90 PMID: 9663684
  18. FACS-optimized mutants of the green fluorescent protein (GFP).
    Gene. 1996;173(1 Spec No):33-8 PMID: 8707053
  19. rRNA transcription rate in Escherichia coli.
    J Bacteriol. 1991 Oct;173(20):6647-9 PMID: 1717439
  20. The complete genome sequence of the gram-positive bacterium Bacillus subtilis.
    Nature. 1997 Nov 20;390(6657):249-56 PMID: 9384377
  21. A vital stain for studying membrane dynamics in bacteria: a novel mechanism controlling septation during Bacillus subtilis sporulation.
    Mol Microbiol. 1999 Feb;31(4):1149-59 PMID: 10096082
  22. The bacterial nucleoid revisited.
    Microbiol Rev. 1994 Jun;58(2):211-32 PMID: 7521510
  23. Bipolar localization of a chromosome partition protein in Bacillus subtilis.
    Proc Natl Acad Sci U S A. 1997 Apr 29;94(9):4721-6 PMID: 9114058
  24. GFP vectors for controlled expression and dual labelling of protein fusions in Bacillus subtilis.
    Gene. 1999 Feb 4;227(1):101-10 PMID: 9931458
  25. Ribosome recycling by ribosome recycling factor (RRF)--an important but overlooked step of protein biosynthesis.
    Adv Biophys. 1996;32:121-201 PMID: 8781287
  26. Identification and characterization of a bacterial chromosome partitioning site.
    Cell. 1998 Mar 6;92(5):675-85 PMID: 9506522
  27. The organization of replication and transcription.
    Science. 1999 Jun 11;284(5421):1790-5 PMID: 10364545
  28. Selection of the midcell division site in Bacillus subtilis through MinD-dependent polar localization and activation of MinC.
    Mol Microbiol. 1999 Jul;33(1):84-96 PMID: 10411726
  29. Visualization of bacterial genes in action.
    Science. 1970 Jul 24;169(3943):392-5 PMID: 4915822
  30. Localization of transcribing genes in the bacterial cell by means of high resolution autoradiography.
    J Mol Biol. 1975 Nov 15;98(4):797-810 PMID: 811810
  31. Association of the histone-like protein HBsu with the nucleoid of Bacillus subtilis.
    J Bacteriol. 1997 Mar;179(6):2060-4 PMID: 9068655
  32. Construction and initial characterization of Escherichia coli strains with few or no intact chromosomal rRNA operons.
    J Bacteriol. 1999 Jun;181(12):3803-9 PMID: 10368156
  33. Growth rate of polypeptide chains as a function of the cell growth rate in a mutant of Escherichia coli 15.
    J Mol Biol. 1971 Feb 14;55(3):563-8 PMID: 4927947
  34. Direct evidence for active segregation of oriC regions of the Bacillus subtilis chromosome and co-localization with the SpoOJ partitioning protein.
    Mol Microbiol. 1997 Sep;25(5):945-54 PMID: 9364919
  35. Metabolic growth rate control in Escherichia coli may be a consequence of subsaturation of the macromolecular biosynthetic apparatus with substrates and catalytic components.
    Microbiol Rev. 1990 Jun;54(2):89-100 PMID: 1694554
  36. Structure and partitioning of bacterial DNA: determined by a balance of compaction and expansion forces?
    FEMS Microbiol Lett. 1995 Sep 15;131(3):235-42 PMID: 7557335
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
2000-02-15
Pages
710-8
Language
English
Region
England
NLM ID
8208664
PMCID
PMC305609
Subset
IM
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]