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

• Published in: International journal of biological macromolecules Vol. 263; p. 130262
• Main Authors: Murugesan, Akshaya, Alshagrawi, Reshod A., Thiyagarajan, Ramesh, Kandhavelu, Meenakshisundaram
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.04.2024
• Subjects: Cell cycle; Noise components; Promoter dynamics; Protein expression; Protein expression; Promoter dynamics; Noise components; Cell cycle
• ISSN: 0141-8130; 1879-0003
• DOI: 10.1016/j.ijbiomac.2024.130262

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. •Dual colour fluorescence protein reporter system detects total cell-to-cell variation.•Total variation increases from Lag to Exponential phase and decreases from Retardation to Stationary phase.•Presence of higher Mg2+ increases population homogeneity than lower Mg2+ concentration.•Intrinsic noise measured from single- and multi-copy plasmids differ by 25%.