A dual fluorescence protein expression system detects cell cycle dependent protein noise

Akshaya Murugesan, Reshod A. Alshagrawi, Ramesh Thiyagarajan, Meenakshisundaram Kandhavelu

Research output: Contribution to journalArticleScientificpeer-review

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Abstract

Inherently identical cells exhibit significant phenotypic variation. It can be essential for many biological processes and is known to arise from stochastic, ‘noisy’, gene expression that is determined by intrinsic and extrinsic components. It is now obvious that the noise varies as a function of inducer concentration. However, its fluctuation over the cell cycle is limited. Applying dual colour fluorescence protein reporter system, Cyan Fluorescent Protein (CFP) and Yellow fluorescent protein (YFP) tagged multi-copy plasmids, we determine variation of the noise components over the phases in lac promoter induced by Isopropyl β-D-1-thiogalactopyranoside (IPTG) and in presence of additional Magnesium, Mg2+ ion. We, also, estimate the how such system deviates from observations of single-copy plasmid. Found 25 % difference between multi-copy system and single-copy system clarifies that observed noise is considerable and estimates population behaviour during the cell cycle. We show that total variation in cells induced with IPTG is determined by higher extrinsic than intrinsic noise. It increases from Lag to Exponential phase and decreases from Retardation to Stationary phase. By observing slow and fast dividing cells, we show that 5 mM Mg2+ increases population homogeneity compared to 2.5 mM Mg2+ in the environment. The experimental data obtained using dual colour fluorescence protein reporter system demonstrates that protein expression noise, depending on intra cellular ionic concentration, is tightly controlled by phase of the cell.

Original languageEnglish
Article number130262
Number of pages7
JournalInternational Journal of Biological Macromolecules
Volume263
DOIs
Publication statusPublished - Apr 2024
Publication typeA1 Journal article-refereed

Keywords

  • Cell cycle
  • Noise components
  • Promoter dynamics
  • Protein expression

Publication forum classification

  • Publication forum level 1

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Molecular Biology

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