Sequential Establishment of Stripe Patterns in an Expanding Cell Population

A synthetic circuit implementing density-controlled bacterial motility autonomously produces a tunable stripe pattern. Periodic stripe patterns are ubiquitous in living organisms, yet the underlying developmental processes are complex and difficult to disentangle. We describe a synthetic genetic circuit that couples cell density and motility. This system enabled programmed Escherichia coli cells to form periodic stripes of high and low cell densities sequentially and autonomously. Theoretical and experimental analyses reveal that the spatial structure arises from a recurrent aggregation process at the front of the continuously expanding cell population. The number of stripes formed could be tuned by modulating the basal expression of a single gene. The results establish motility control as a simple route to establishing recurrent structures without requiring an extrinsic pacemaker.

[1]  R. Fisher.  THE WAVE OF ADVANCE OF ADVANTAGEOUS GENES , 1937 .

[2]  A. Turing.  The chemical basis of morphogenesis , 1952, Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences.

[3]  J. Adler.  Chemotaxis in Bacteria , 1966, Science.

[4]  Richard Gran,et al.  On the Convergence of Random Search Algorithms In Continuous Time with Applications to Adaptive Control , 1970, IEEE Trans. Syst. Man Cybern..

[5]  H. Berg.  Chemotaxis in bacteria. , 1975, Annual review of biophysics and bioengineering.

[6]  N. Otsu.  A threshold selection method from gray level histograms , 1979 .

[7]  C. Nicholson,et al.  Ion diffusion modified by tortuosity and volume fraction in the extracellular microenvironment of the rat cerebellum. , 1981, The Journal of physiology.

[8]  J. S. Parkinson,et al.  Isolation and behavior of Escherichia coli deletion mutants lacking chemotaxis functions , 1982, Journal of bacteriology.

[9]  H. Berg,et al.  Migration of bacteria in semisolid agar. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[10]  H. Berg,et al.  Complex patterns formed by motile cells of Escherichia coli , 1991, Nature.

[11]  R. M. Ford,et al.  Measurement of bacterial random motility and chemotaxis coefficients: I. Stopped‐flow diffusion chamber assay , 1991, Biotechnology and bioengineering.

[12]  H. Fujikawa.  Periodic growth of Bacillus subtilis colonies on agar plates , 1992 .

[13]  Charles Nicholson,et al.  Ion-selective microelectrodes and diffusion measurements as tools to explore the brain cell microenvironment , 1993, Journal of Neuroscience Methods.

[14]  J. A. Quinn,et al.  Random motility of swimming bacteria: Single cells compared to cell populations , 1994 .

[15]  H. Berg,et al.  Spatio-temporal patterns generated by Salmonella typhimurium. , 1995, Biophysical journal.

[16]  M. Simon,et al.  In vivo and in vitro characterization of Escherichia coli protein CheZ gain- and loss-of-function mutants , 1996, Journal of bacteriology.

[17]  Frank Buchholz,et al.  A new logic for DNA engineering using recombination in Escherichia coli , 1998, Nature Genetics.

[18]  L. Poulsen,et al.  New Unstable Variants of Green Fluorescent Protein for Studies of Transient Gene Expression in Bacteria , 1998, Applied and Environmental Microbiology.

[19]  Herbert Levine,et al.  Cooperative self-organization of microorganisms , 2000 .

[20]  M. Matsushita,et al.  Dynamic Aspects of the Structured Cell Population in a Swarming Colony of Proteus mirabilis , 2000, Journal of bacteriology.

[21]  E. Greenberg,et al.  Detection, purification, and structural elucidation of the acylhomoserine lactone inducer of Vibrio fischeri luminescence and other related molecules. , 2000, Methods in enzymology.

[22]  R M Ford,et al.  Quantification of random motility and chemotaxis bacterial transport coefficients using individual-cell and population-scale assays. , 2001, Biotechnology and bioengineering.

[23]  C. Nicholson,et al.  Quantitative dual‐probe microdialysis: evaluation of [3H]mannitol diffusion in agar and rat striatum , 2002, Journal of neurochemistry.

[24]  Steve Atkinson,et al.  N-Acylhomoserine Lactones Undergo Lactonolysis in a pH-, Temperature-, and Acyl Chain Length-Dependent Manner during Growth of Yersinia pseudotuberculosis and Pseudomonas aeruginosa , 2002, Infection and Immunity.

[25]  O. Pourquié.  The Segmentation Clock: Converting Embryonic Time into Spatial Pattern , 2003, Science.

[26]  L. Niswander.  Pattern formation: old models out on a limb , 2003, Nature Reviews Genetics.

[27]  K. Uematsu,et al.  Inhibition of Wnt-2-mediated signaling induces programmed cell death in non-small-cell lung cancer cells , 2004, Oncogene.

[28]  M. Radmacher,et al.  pH Regulates Genes for Flagellar Motility, Catabolism, and Oxidative Stress in Escherichia coli K-12 , 2005, Journal of bacteriology.

[29]  M. Isalan,et al.  Engineering Gene Networks to Emulate Drosophila Embryonic Pattern Formation , 2005, PLoS biology.

[30]  Nathan C Shaner,et al.  A guide to choosing fluorescent proteins , 2005, Nature Methods.

[31]  R. Weiss,et al.  Ultrasensitivity and noise propagation in a synthetic transcriptional cascade. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[32]  S. Basu,et al.  A synthetic multicellular system for programmed pattern formation , 2005, Nature.

[33]  B. Bassler,et al.  Quorum sensing: cell-to-cell communication in bacteria. , 2005, Annual review of cell and developmental biology.

[34]  N. Costantino,et al.  A set of recombineering plasmids for gram-negative bacteria. , 2006, Gene.

[35]  E. Davidson,et al.  Gene Regulatory Networks and the Evolution of Animal Body Plans , 2006, Science.

[36]  D. Montell.  The social lives of migrating cells in Drosophila. , 2006, Current opinion in genetics & development.

[37]  T. Terwilliger,et al.  Engineering and characterization of a superfolder green fluorescent protein , 2006, Nature Biotechnology.

[38]  G. Sezonov,et al.  Escherichia coli Physiology in Luria-Bertani Broth , 2007, Journal of bacteriology.

[39]  Jing Wang,et al.  Visualizing the proteome of Escherichia coli: an efficient and versatile method for labeling chromosomal coding DNA sequences (CDSs) with fluorescent protein genes , 2007, Nucleic acids research.

[40]  Saeed Tavazoie,et al.  A Comprehensive Genetic Characterization of Bacterial Motility , 2007, PLoS genetics.

[41]  P. Rørth.  Collective guidance of collective cell migration. , 2007, Trends in cell biology.

[42]  Markus Affolter,et al.  The Decapentaplegic morphogen gradient: from pattern formation to growth regulation , 2007, Nature Reviews Genetics.

[43]  Tom L Richard,et al.  Engineering motility as a phenotypic response to LuxI/R-dependent quorum sensing in Escherichia coli. , 2008, Biotechnology and bioengineering.

[44]  Seungyong Lee,et al.  Real-time PCR determination of rRNA gene copy number: absolute and relative quantification assays with Escherichia coli , 2008, Applied Microbiology and Biotechnology.

[45]  M E Cates,et al.  Statistical mechanics of interacting run-and-tumble bacteria. , 2008, Physical review letters.

[46]  T. Hwa,et al.  Growth Rate-Dependent Global Effects on Gene Expression in Bacteria , 2009, Cell.

[47]  N. Thompson,et al.  Quantifying green fluorescent protein diffusion in Escherichia coli by using continuous photobleaching with evanescent illumination. , 2009, The journal of physical chemistry. B.

[48]  C. Chuong,et al.  Pattern formation today. , 2009, The International journal of developmental biology.

[49]  Shankar Mukherji,et al.  Synthetic biology: understanding biological design from synthetic circuits , 2009, Nature Reviews Genetics.

[50]  M E Cates,et al.  Arrested phase separation in reproducing bacteria creates a generic route to pattern formation , 2010, Proceedings of the National Academy of Sciences.

[51]  A. Aman,et al.  Cell migration during morphogenesis. , 2010, Developmental biology.

[52]  Ahmad S. Khalil,et al.  Synthetic biology: applications come of age , 2010, Nature Reviews Genetics.

[53]  M. Elowitz,et al.  Build life to understand it , 2010, Nature.