Effects of growth rate and promoter activity on single-cell protein expression

Protein expression in a single cell depends on its global physiological state. Moreover, genetically-identical cells exhibit variability (noise) in protein expression, arising from the stochastic nature of biochemical processes, cell growth and division. While it is well understood how cellular growth rate influences mean protein expression, little is known about the relationship between growth rate and noise in protein expression. Here we quantify this relationship in Bacillus subtilis by a novel combination of experiments and theory. We measure the effects of promoter activity and growth rate on the expression of a fluorescent protein in single cells. We disentangle the observed protein expression noise into protein-specific and systemic contributions, using theory and variance decomposition. We find that noise in protein expression depends solely on mean expression levels, regardless of whether expression is set by promoter activity or growth rate, and that noise increases linearly with growth rate. Our results can aid studies of (synthetic) gene circuits of single cells and their condition dependence.

[1]  Miguel Rocha,et al.  Transcript level and sequence determinants of protein abundance and noise in Escherichia coli , 2014, Nucleic acids research.

[2]  M. Richmond,et al.  Rate of growth of Bacillus cereus between divisions. , 1962, Journal of general microbiology.

[3]  N. Friedman,et al.  Stochastic protein expression in individual cells at the single molecule level , 2006, Nature.

[4]  P. Dennis,et al.  Macromolecular Composition During Steady-State Growth of Escherichia coli B/r , 1974, Journal of bacteriology.

[5]  Johan Paulsson,et al.  Separating intrinsic from extrinsic fluctuations in dynamic biological systems , 2011, Proceedings of the National Academy of Sciences.

[6]  E. van Nimwegen,et al.  Figures and figure supplements Expression noise facilitates the evolution of gene regulation , 2015 .

[7]  Jatin Narula,et al.  Slowdown of growth controls cellular differentiation , 2016, Molecular systems biology.

[8]  Albert Siryaporn,et al.  Superresolution imaging of ribosomes and RNA polymerase in live Escherichia coli cells , 2012, Molecular microbiology.

[9]  R. Milo,et al.  Promoters maintain their relative activity levels under different growth conditions , 2013, Molecular systems biology.

[10]  Nam Ki Lee,et al.  Contribution of RNA polymerase concentration variation to protein expression noise , 2014, Nature Communications.

[11]  V. Shahrezaei,et al.  Connecting growth with gene expression: of noise and numbers. , 2015, Current opinion in microbiology.

[12]  B. Schwikowski,et al.  Condition-Dependent Transcriptome Reveals High-Level Regulatory Architecture in Bacillus subtilis , 2012, Science.

[13]  E. Yarovaya,et al.  Effect of fibrinogen on platelet reactivity measured by the VerifyNow P2Y12 assay , 2016, Biochemistry (Moscow).

[14]  R. Milo,et al.  Protein Dynamics in Individual Human Cells: Experiment and Theory , 2009, PloS one.

[15]  F. Bruggeman,et al.  The volumes and transcript counts of single cells reveal concentration homeostasis and capture biological noise , 2015, Molecular biology of the cell.

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

[17]  Michael L. Simpson,et al.  The Low Noise Limit in Gene Expression , 2015, PloS one.

[18]  Jeffrey W. Smith,et al.  Stochastic Gene Expression in a Single Cell , 2022 .

[19]  J. Paulsson Summing up the noise in gene networks , 2004, Nature.

[20]  A. Kruse,et al.  SEDS proteins are a widespread family of bacterial cell wall polymerases , 2016, Nature.

[21]  John T. Sauls,et al.  Cell-Size Control and Homeostasis in Bacteria , 2015, Current Biology.

[22]  A. Grossman,et al.  In Vivo Effects of Sporulation Kinases on Mutant Spo0A Proteins in Bacillus subtilis , 2001, Journal of bacteriology.

[23]  Johan Paulsson,et al.  Random partitioning of molecules at cell division , 2011, Proceedings of the National Academy of Sciences.

[24]  S. Klumpp,et al.  Dilution and the theoretical description of growth-rate dependent gene expression , 2013, Journal of biological engineering.

[25]  Ertugrul M. Ozbudak,et al.  Regulation of noise in the expression of a single gene , 2002, Nature Genetics.

[26]  Terence Hwa,et al.  Bacterial growth: global effects on gene expression, growth feedback and proteome partition. , 2014, Current opinion in biotechnology.

[27]  K. Hellingwerf,et al.  Effects of Phosphorelay Perturbations on Architecture, Sporulation, and Spore Resistance in Biofilms of Bacillus subtilis , 2006, Journal of bacteriology.

[28]  K. Gerdes,et al.  RETRACTED: (p)ppGpp Controls Bacterial Persistence by Stochastic Induction of Toxin-Antitoxin Activity , 2013, Cell.

[29]  P. Swain,et al.  Intrinsic and extrinsic contributions to stochasticity in gene expression , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[30]  J. Derisi,et al.  Single-cell proteomic analysis of S. cerevisiae reveals the architecture of biological noise , 2006, Nature.

[31]  David H Burkhardt,et al.  Quantifying Absolute Protein Synthesis Rates Reveals Principles Underlying Allocation of Cellular Resources , 2014, Cell.

[32]  F. Bruggeman,et al.  Contributions of cell growth and biochemical reactions to nongenetic variability of cells. , 2014, Biophysical journal.

[33]  J. Helmann,et al.  Manganese homeostasis in Bacillus subtilis is regulated by MntR, a bifunctional regulator related to the diphtheria toxin repressor family of proteins , 2000, Molecular microbiology.

[34]  E. Cox,et al.  Real-Time Kinetics of Gene Activity in Individual Bacteria , 2005, Cell.

[35]  Matthias Heinemann,et al.  Condition-Dependent Cell Volume and Concentration of Escherichia coli to Facilitate Data Conversion for Systems Biology Modeling , 2011, PloS one.

[36]  J. Bell,et al.  Experiment and Theory , 1968 .

[37]  N. Grover,et al.  On microbial states of growth † , 1995, Molecular microbiology.

[38]  F. Bruggeman,et al.  Single yeast cells vary in transcription activity not in delay time after a metabolic shift , 2014, Nature Communications.

[39]  Ido Golding,et al.  Genetic Determinants and Cellular Constraints in Noisy Gene Expression , 2013, Science.

[40]  P. Swain,et al.  Gene Regulation at the Single-Cell Level , 2005, Science.

[41]  D. J. Kiviet,et al.  Stochasticity of metabolism and growth at the single-cell level , 2014, Nature.

[42]  A. van Oudenaarden,et al.  Noise Propagation in Gene Networks , 2005, Science.

[43]  D. Henner,et al.  Use of the Escherichia coli lac repressor and operator to control gene expression in Bacillus subtilis. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[44]  T. Hwa,et al.  Interdependence of Cell Growth and Gene Expression: Origins and Consequences , 2010, Science.

[45]  Johan Paulsson,et al.  Models of stochastic gene expression , 2005 .

[46]  Nir Friedman,et al.  Linking stochastic dynamics to population distribution: an analytical framework of gene expression. , 2006, Physical review letters.

[47]  Paul J. Choi,et al.  Quantifying E. coli Proteome and Transcriptome with Single-Molecule Sensitivity in Single Cells , 2010, Science.

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

[49]  Reinhard Wolf,et al.  Coding-Sequence Determinants of Gene Expression in Escherichia coli , 2009 .

[50]  Noise and Numbers , 2006 .