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Related Experiment Video

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Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay

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Published on: May 3, 2018

A phosphorylation-controlled switch confers cell cycle-dependent protein relocalization.

Xiaofu Cao^1,2, Shiying Huang^1,2, Mateusz M Wagner^2,3

¹Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York, United States, 14853.

Biorxiv : the Preprint Server for Biology

|June 19, 2024

Summary

This summary is machine-generated.

Researchers developed a novel system called MARS (Mitosis-enabled Anchor-away/Recruiter System) to precisely control protein localization during cell division without external triggers. This tool aids in studying mitosis and editing cell membranes.

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Area of Science:

Cell Biology
Molecular Biology
Biochemistry

Background:

Studying protein function during mitosis is crucial but challenging due to cell sensitivity.
Existing tools for protein manipulation often require external triggers, which can disrupt delicate cellular processes like mitosis.

Purpose of the Study:

To develop a novel, stimulus-independent system for precise protein recruitment during mitosis.
To enable the study of spatiotemporally defined protein functions during cell division.

Main Methods:

Exploited a naturally occurring, cell cycle-dependent localization change of the PLEKHA5 protein.
Engineered a 15-kDa module from PLEKHA5 to create the Mitosis-enabled Anchor-away/Recruiter System (MARS).
Utilized MARS for direct fusion or GFP-nanobody interactions to recruit protein cargoes.

Main Results:

MARS enables mitosis-specific protein recruitment to the plasma membrane without exogenous stimuli.
Demonstrated MARS application for 'knock sideways' experiments to displace proteins during mitosis.
Showcased conditional recruitment of enzymes for mitosis-selective lipid editing of the plasma membrane.

Conclusions:

MARS provides a powerful, non-perturbative tool for investigating mitosis.
This system overcomes limitations of exogenous triggers in studying cell division.
MARS facilitates new approaches for functional genomics and cell membrane engineering during mitosis.