Related Experiment Video
Updated: Aug 15, 2025
10:02
Applications of pHluorin for Quantitative, Kinetic and High-throughput Analysis of Endocytosis in Budding Yeast
Published on: October 23, 2016
11.0K
Using Budding Yeast as a Model to Understand Dynein-Mediated Cargo Transport
Kari H Ecklund1, Matthew G Marzo1, Steven M Markus2
1Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO, USA.
Methods in Molecular Biology (Clifton, N.J.)
|January 5, 2023
Summary
Researchers developed protocols to measure cytoplasmic dynein-1 motor protein activity in budding yeast. This system allows quantitative analysis of dynein-mediated spindle movements, aiding the study of this essential motor protein.
Area of Science:
- Cell Biology
- Molecular Motors
- Biophysics
Background:
- Cytoplasmic dynein-1 is a crucial minus end-directed microtubule motor protein.
- Its regulation is complex, involving intrinsic and extrinsic factors.
- Studying dynein in higher eukaryotes is challenging due to its diverse functions.
Purpose of the Study:
- To establish a simplified model system for studying dynein activity.
- To provide detailed protocols for quantitative measurement of dynein function.
- To enable assessment of mutations affecting dynein or its regulators.
Main Methods:
- Utilizing budding yeast as a model organism, where dynein has a single known function.
- Employing live-cell fluorescence microscopy to visualize cellular processes.
- Applying computational methods for tracking and quantifying dynein-mediated spindle movements.
Main Results:
- Developed and detailed protocols for quantitative measurement of dynein activity.
- Established a robust system for analyzing dynein-mediated spindle positioning.
- Demonstrated the utility of budding yeast for dissecting motor protein function.
Conclusions:
- Budding yeast offers a powerful and simplified system for studying conserved motor proteins like dynein.
- The provided protocols enable quantitative assessment of dynein motility parameters.
- These methods are broadly applicable for fluorescence microscopy studies in budding yeast.
Related Concept Videos
01:43The Movement of Organelles and Vesicles
4.6K
In eukaryotic cells, cytoskeletal filaments such as actin, microtubules, and intermediate filaments form a mesh-like cytoskeletal network. These filaments serve as tracks for transporting cellular cargo. Specialized motor proteins use the chemical energy stored in adenosine triphosphate (ATP) for this transport. During interphase, microtubules are polarized, with the plus-end towards the cell periphery and the minus-end towards the cell center. Two microtubule-associated motor proteins,...
4.6K
01:32Microtubule Associated Motor Proteins
8.3K
Eukaryotic cells have different motor proteins for transporting various cargo within the cell. These motor proteins differ based on the filament they associate with, the direction they move within the cell, and the type of cargo they transport. Motor proteins that associate with microtubules are known as microtubule-associated motor proteins. There are two families of microtubule-associated motor proteins —Kinesins and Dyneins. Both these proteins assist in the transport of cellular...
8.3K
01:32Pinching-off of Coated Vesicles
3.2K
Vesicle budding is orchestrated by distinct cytosolic proteins such as adaptor proteins, coat proteins, and GTPases. To initiate vesicle budding, membrane-bending proteins containing crescent-shaped BAR domains bind to the lipid heads in the bilayer and distort the membrane to form a protein-coated vesicle bud. Adaptors proteins such as AP2 for clathrin-coated vesicles can nucleate on the deformed membrane. Finally, coat proteins such as clathrin or COPI and COPII assemble into a coat forming...
3.2K
01:12Clathrin Coated Vesicles
7.1K
Clathrin-coated vesicles use endocytosis to transport receptors and lysosomal hydrolases from the Golgi to the lysosome in the late secretory pathway. Clathrin-mediated endocytosis was the first described endocytic process, and Clathrin-coated vesicles remain one of the most well-studied transport vesicles. The molecular machinery that generates clathrin-coated vesicles comprises over 50 proteins that precisely coordinate vesicle formation. Cell surface receptors concentrated in indented sites...
7.1K
01:10Intracellular Movement of Viruses and Bacteria
2.9K
Intracellular bacteria and viruses often comprise a group of highly infectious pathogens that can cause several diseases. Bacterial pathogens include those belonging to the genus Rickettsia responsible for conditions such as rocky mountain spotted fever and the Mediterranean spotted fever; Chlamydia, a genus responsible for a sexually transmitted disease; Coxiella burnetii, an agent responsible for Q fever. Viral pathogens include vaccinia—a poxvirus, and herpes simplex virus—a...
2.9K
01:24Microtubules in Cell Motility
3.4K
Microtubules are thick hollow cylindrical proteins that help form the cytoskeleton. Microtubules have varied roles in the cell. These filaments help form cellular appendages like cilia and flagella, which are responsible for locomotion. The cilia arise from basal bodies, separated from the main body by a membrane-like structure forming the transition zone. This zone is the gate for the entry of lipids and proteins, creating a unique composition of lipids and proteins in the ciliary membrane and...
3.4K