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

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.