Nucleotide Pool Dynamics
Nucleotide Pool Dynamics - Related Videos
02:43
Thin-Layer Chromatography-Based Separation of (p)ppGpp Nucleotides Isolated from Stress-Induced Bacteria
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2025
Source: Fernández-Coll, L. and Cashel, M. Using Microtiter Dish Radiolabeling for Multiple In Vivo Measurements Of Escherichia coli (p)ppGpp Followed by Thin Layer Chromatography. J. Vis. Exp. (2019)This video presents a high-throughput protocol for measuring the levels and dynamics of the bacterial stress-response nucleotides ppGpp and pppGpp using thin-layer chromatography (TLC). The method provides a powerful, rapid, and cost-effective approach to quantifying (p)ppGpp in bacterial cultures...
14:06
Detection of Rare Genomic Variants from Pooled Sequencing Using SPLINTER
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Cited by 10 •
2012
Pooled DNA sequencing is a fast and cost-effective strategy to detect rare variants associated with complex phenotypes in large cohorts. Here we describe the computational analysis of pooled, next-generation sequencing of 32 cancer-related genes using the SPLINTER software package. This method is scalable, and applicable to any phenotype of interest.
09:12
MISSION LentiPlex Pooled shRNA Library Screening in Mammalian Cells
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Cited by 11 •
2011
Here we use a human LentiPlex pooled library and traditional sequencing methods to identify gene targets promoting cell survival. We demonstrate how to set up and deconvolute a LentiPlex screen and validate the results.
06:38
Plant Sample Preparation for Nucleoside/Nucleotide Content Measurement with An HPLC-MS/MS
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Cited by 3 •
2021
A precise and reproducible method for in vivo nucleosides/nucleotides quantification in plants is described here. This method employs an HPLC-MS/MS.
07:32
Pool-Boiling Heat-Transfer Enhancement on Cylindrical Surfaces with Hybrid Wettable Patterns
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Cited by 17 •
2017
Pool-boiling heat-transfer experiments were carried out to observe the effects of hybrid wettable patterns on the heat-transfer coefficient (HTC). The parameters of investigation are the number of interlines and the pattern orientation of the modified wettable surface.
FAQs
Cells regulate nucleotide availability through several connected routes rather than a single pathway. De novo synthesis produces nucleotides from metabolic precursors, salvage recycles available components, phosphorylation changes their usable forms, and degradation removes excess or damaged metabolites. Coordinated transport between compartments further distributes these molecules, helping match local supply with RNA production, DNA replication, and signaling demands.
DNA replication requires an appropriate supply and composition of deoxyribonucleotides, while ribonucleotides support RNA synthesis and other cellular functions. If pool size or composition becomes abnormal, replication can stall and repair processes can be impaired. These disruptions may also increase mutagenesis, linking nucleotide regulation directly to genome stability and the fidelity of genetic information.
Feedback control adjusts pathway activity according to growth, nutrient availability, and DNA replication demand. Increased cellular activity can alter the need for nucleotide production, whereas limited nutrients can constrain synthesis and change the balance among production, recycling, and breakdown. This responsive regulation prevents pathway activity from becoming disconnected from the cell's immediate metabolic and replication requirements.
Transport between cellular compartments helps place nucleotide metabolites where they are needed and influences their local abundance and turnover. Consequently, total cellular measurements may not fully describe the conditions experienced by replication, transcription, or signaling processes in a particular compartment. Considering transport provides a more precise view of how metabolic regulation supports distinct cellular activities.
Measuring changes in nucleotide abundance, composition, and turnover allows researchers to connect metabolic regulation with genome stability. Such analyses can reveal whether altered pathway activity accompanies replication problems, impaired repair, or increased mutagenesis. The resulting information helps distinguish changes in nucleotide metabolism from their effects on genetic maintenance and cellular function.
These fields depend on understanding how altered nucleotide regulation affects cellular growth and genetic stability. In cancer biology, disrupted pools can expose metabolic dependencies; in antimicrobial research, nucleotide pathways provide relevant biological targets. Drug-development studies can use pool changes to identify therapeutic vulnerabilities and evaluate whether pathway perturbation produces replication or repair defects.