NHSL3 controls single and collective cell migration through two distinct mechanisms.

Area of Science:

• Cell Biology
• Molecular Mechanisms
• Genetics

Background:

• The molecular underpinnings of cell migration are not fully elucidated.
• Nance-Horan Syndrome Like 3 (NHSL3) is a recently identified gene family member.

Purpose of the Study:

• To investigate the role of NHSL3 in single-cell and collective cell migration.
• To identify specific NHSL3 isoforms and their protein interactions critical for distinct migration behaviors.

Main Methods:

• Utilized knock-out cell models to assess migration persistence and follower cell behavior.
• Employed proteomics to determine the partner repertoire of NHSL3 isoforms.
• Applied an AlphaFold2-based pipeline for predicting protein-protein interactions and binding sites.
• Conducted rescue experiments with specific NHSL3 isoforms and mutated binding sites.

Main Results:

• NHSL3 knock-out cells exhibited increased persistence in single-cell migration but impaired follower cell function in collective migration.
• Distinct NHSL3 isoforms interact with different protein partners: long isoforms with MENA/VASP at cell junctions, and short isoforms with 14-3-3θ in lamellipodia.
• Mutations in critical binding sites abolished the rescue effects, confirming the functional importance of these interactions.

Conclusions:

• NHSL3 regulates single-cell migration via interaction with 14-3-3θ in lamellipodia.
• NHSL3 regulates collective cell migration through interaction with MENA/VASP proteins at cell-cell junctions.
• NHSL3 employs distinct molecular mechanisms to control different modes of cell migration.