Related Experiment Video

Updated: Jul 13, 2026

Using Caenorhabditis elegans as a Model System to Study Protein Homeostasis in a Multicellular Organism
12:38

Using Caenorhabditis elegans as a Model System to Study Protein Homeostasis in a Multicellular Organism

Published on: December 18, 2013

Counting cytokinesis proteins globally and locally in fission yeast

Jian-Qiu Wu1, Thomas D Pollard

  • 1Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520-8103, USA.

Science (New York, N.Y.)
|October 15, 2005
PubMed

Insights

Researchers measured protein concentrations in fission yeast using yellow fluorescent protein (YFP) fusions. This microscopy method quantifies global and local protein levels, revealing significant cellular compartmentalization.

Area of Science:

  • Cell biology
  • Biophysics
  • Molecular biology

Background:

  • Understanding protein concentration is crucial for cell function.
  • Previous methods had limitations in measuring global and local protein levels simultaneously.

Purpose of the Study:

  • To develop and validate a method for measuring global and local protein concentrations in vivo.
  • To quantify the concentrations of 28 cytoskeletal and signaling proteins in Schizosaccharomyces pombe.

Main Methods:

  • Utilized fluorescence microscopy to measure yellow fluorescent protein (YFP) fusion protein concentrations.
  • Employed native promoters for controlled expression of functional YFP fusion proteins.
  • Validated fluorescence measurements against quantitative immunoblotting for protein concentration accuracy.

Main Results:

  • Established a direct proportionality between fluorescence and protein concentration.
  • Determined global cytoplasmic concentrations for 28 proteins, ranging from 0.04 to 63 micromolar.
  • Observed significant protein enrichment (up to 7500-fold) in specific cellular compartments like contractile rings and spindle pole bodies during the cell cycle.

Conclusions:

  • The developed fluorescence microscopy approach accurately measures global and local protein concentrations.
  • Demonstrated substantial spatial and temporal regulation of protein abundance in fission yeast.
  • This method is applicable for quantifying any fusion protein's concentration in vivo.

More Related Videos

Combining Magnetic Sorting of Mother Cells and Fluctuation Tests to Analyze Genome Instability During Mitotic Cell Aging in Saccharomyces cerevisiae11:08

Combining Magnetic Sorting of Mother Cells and Fluctuation Tests to Analyze Genome Instability During Mitotic Cell Aging in Saccharomyces cerevisiae

Published on: October 16, 2014

Manipulation and Analysis of Cell Cycle-Dependent Processes in Budding Yeast08:13

Manipulation and Analysis of Cell Cycle-Dependent Processes in Budding Yeast

Published on: September 26, 2025

Related Concept Videos

Protein and Protein Structure02:15

Protein and Protein Structure

Proteins are one of the most abundant organic molecules in living systems and have the most diverse range of functions of all macromolecules. Proteins may be structural, regulatory, contractile, or protective. They may serve in transport, storage, or membranes; or they may be toxins or enzymes. Their structures, like their functions, vary greatly. They are all, however, amino acid polymers arranged in a linear sequence.
A protein's shape is critical to its function. For example, an enzyme can...
Proteins: From Genes to Degradation02:11

Proteins: From Genes to Degradation

Within a biological system, the DNA encodes the RNA, and the nucleotide sequence in the RNA further defines the amino acid sequence in the protein. This is referred to as “The Central Dogma of Molecular Biology” - a term coined by Francis Crick.  Central dogma is a firm principle in biology that defines the flow of genetic information within any life form. The two fundamental steps in central dogma are - transcription and translation.
Transcription is the synthesis of RNA molecules by RNA...
Distribution of Cytoplasmic Content02:33

Distribution of Cytoplasmic Content

Cytokinesis segregates a cell’s chromosomes and organelles into its daughter cells. Organelles divide and grow prior to cell division but cannot be synthesized de novo; therefore, cells must receive at least one copy of each organelle to survive. Currently, many of the details of how the organelles are distributed are not yet fully elucidated.
Distribution of cytoplasmic determinants
The cytoplasm contains various organelles, as well as salts, proteins, and water. The distribution of small...
Distribution of Cytoplasmic Content02:33

Distribution of Cytoplasmic Content

Cytokinesis segregates a cell’s chromosomes and organelles into its daughter cells. Organelles divide and grow prior to cell division but cannot be synthesized de novo; therefore, cells must receive at least one copy of each organelle to survive. Currently, many of the details of how the organelles are distributed are not yet fully elucidated.
Distribution of cytoplasmic determinants
The cytoplasm contains various organelles, as well as salts, proteins, and water. The distribution of small...
Bacterial Protein Maturation01:26

Bacterial Protein Maturation

Bacterial protein maturation is a tightly regulated process that ensures newly synthesized polypeptides achieve correct functional conformations. This maturation involves a series of modifications, folding events, and quality control steps, often assisted by specialized chaperone proteins.N-Terminal ModificationsThe maturation of bacterial polypeptides begins cotranslationally as the polypeptide exits the ribosome. The first amino acid, N-formylmethionine (fMet), is typically modified at the...
Microbes in Food Production01:29

Microbes in Food Production

Microbial fermentation is central to food biotechnology, enhancing flavor, texture, preservation, and stability. Fermentative microorganisms metabolize carbohydrates into organic acids, alcohols, and other metabolites that inhibit spoilage organisms and improve digestibility while contributing distinctive sensory qualities.In baking, amylases naturally present in flour hydrolyze starch into monosaccharides such as glucose, which Saccharomyces cerevisiae ferments anaerobically. Through...