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      Spatial organization of the flow of genetic information in bacteria

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          Abstract

          Eukaryotic cells spatially organize mRNA processes such as translation and mRNA decay. Much less is clear in bacterial cells where the spatial distribution of mature mRNA remains ambiguous. Using a sensitive, quantitative fluorescence in situ hybridization based-method, we show here that in Caulobacter crescentus and Escherichia coli, chromosomally-expressed mRNAs largely display limited dispersion from their site of transcription during their lifetime. We estimate apparent diffusion coefficients at least 2 orders of magnitude lower than expected for freely diffusing mRNA, and provide evidence in C. crescentus that this mRNA localization restricts ribosomal mobility. Furthermore, C. crescentus RNase E appears associated with the DNA independently of its mRNA substrates. Collectively, our findings reveal that bacteria can spatially organize translation and potentially mRNA decay by using the chromosome layout as a template. This chromosome-centric organization has important implications for cellular physiology and for our understanding of gene expression in bacteria.

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          Most cited references32

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          Global analysis of mRNA decay and abundance in Escherichia coli at single-gene resolution using two-color fluorescent DNA microarrays.

          Much of the information available about factors that affect mRNA decay in Escherichia coli, and by inference in other bacteria, has been gleaned from study of less than 25 of the approximately 4,300 predicted E. coli messages. To investigate these factors more broadly, we examined the half-lives and steady-state abundance of known and predicted E. coli mRNAs at single-gene resolution by using two-color fluorescent DNA microarrays. An rRNA-based strategy for normalization of microarray data was developed to permit quantitation of mRNA decay after transcriptional arrest by rifampicin. We found that globally, mRNA half-lives were similar in nutrient-rich media and defined media in which the generation time was approximately tripled. A wide range of stabilities was observed for individual mRNAs of E. coli, although approximately 80% of all mRNAs had half-lives between 3 and 8 min. Genes having biologically related metabolic functions were commonly observed to have similar stabilities. Whereas the half-lives of a limited number of mRNAs correlated positively with their abundance, we found that overall, increased mRNA stability is not predictive of increased abundance. Neither the density of putative sites of cleavage by RNase E, which is believed to initiate mRNA decay in E. coli, nor the free energy of folding of 5' or 3' untranslated region sequences was predictive of mRNA half-life. Our results identify previously unsuspected features of mRNA decay at a global level and also indicate that generalizations about decay derived from the study of individual gene transcripts may have limited applicability.
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            Noise in gene expression determines cell fate in Bacillus subtilis.

            Random cell-to-cell variations in gene expression within an isogenic population can lead to transitions between alternative states of gene expression. Little is known about how these variations (noise) in natural systems affect such transitions. In Bacillus subtilis, noise in ComK, the protein that regulates competence for DNA uptake, is thought to cause cells to transition to the competent state in which genes encoding DNA uptake proteins are expressed. We demonstrate that noise in comK expression selects cells for competence and that experimental reduction of this noise decreases the number of competent cells. We also show that transitions are limited temporally by a reduction in comK transcription. These results illustrate how such stochastic transitions are regulated in a natural system and suggest that noise characteristics are subject to evolutionary forces.
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              Lost in translation: the influence of ribosomes on bacterial mRNA decay.

              The lifetimes of bacterial mRNAs are strongly affected by their association with ribosomes. Events occurring at any stage during translation, including ribosome binding, polypeptide elongation, or translation termination, can influence the susceptibility of mRNA to ribonuclease attack. Ribosomes usually act as protective barriers that impede mRNA cleavage, but in some instances they can instead trigger the decay of the mRNA to which they are bound or send a signal that leads to widespread mRNA destabilization within a cell. The influence of translation on mRNA decay provides a quality-control mechanism for minimizing the use of poorly or improperly translated mRNAs as templates for the production of abnormal proteins that might be toxic to bacteria.
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                Author and article information

                Journal
                0410462
                6011
                Nature
                Nature
                0028-0836
                1476-4687
                17 May 2010
                20 June 2010
                1 July 2010
                1 January 2011
                : 466
                : 7302
                : 77-81
                Affiliations
                [1 ]Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06520, USA.
                [2 ]Howard Hughes Medical Institute, Yale University, New Haven, CT 06520, USA.
                [3 ]Department of Physics, Yale University, New Haven, CT 06520, USA.
                [4 ]Section of Microbial Pathogenesis, Yale School of Medicine, New Haven, CT 06510, USA.
                Author notes
                [*]

                Equally contributed

                [$]

                Current address: Cutaneous Biology Research Center, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129, USA

                []For correspondence: christine.jacobs-wagner@ 123456yale.edu .

                Author Contributions C.J.W., P.M.L., and A.F.J designed experiments. P.M.L performed the FISH, ribosome and RNaseE experiments, and analyzed FISH and FRAP data. A.F.J. carried out the MS2 experiments and analyzed the data. P.M.L. and J.H. performed the real-time PCR measurements. O.S. developed the tools for image and data analysis. I.S. described and implemented the mathematical model for the analysis of mRNA diffusion. T.E. provided conceptual and data analysis advice. C.J.W, P.M.L. and A.F.J. wrote the paper.

                Article
                nihpa200559
                10.1038/nature09152
                2896451
                20562858
                1f26f825-41d3-4596-852d-16ebed15341c

                Users may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms

                History
                Funding
                Funded by: National Institute of General Medical Sciences : NIGMS
                Award ID: R01 GM065835-07 ||GM
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