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Quantitative Immunofluorescence Assay to Measure the Variation in Protein Levels at Centrosomes
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A role for Cep70 in centriole amplification in multiciliated cells
Sun K Kim1, Eva Brotslaw1, Virginie Thome2
1Department of Cell and Developmental Biology, Lurie Comprehensive Cancer Center, Northwestern University, Feinberg School of Medicine, France.
Developmental Biology
|December 7, 2020
Summary
Centriole amplification in multiciliated cells involves the deuterosome. Researchers found Cep70 protein associates with deuterosomes, regulating centriole biogenesis and formation.
Area of Science:
- Cell Biology
- Developmental Biology
- Molecular Biology
Background:
- Multiciliated cells (MCCs) amplify centrioles via a pseudo-cell cycle.
- Deuterosomes are poorly understood structures involved in this process.
Purpose of the Study:
- To identify novel proteins associated with deuterosomes.
- To investigate the role of Cep70 in centriole amplification and MCC development.
Main Methods:
- Immunofluorescence microscopy in Xenopus embryos.
- Co-immunoprecipitation assays.
- Morpholino-mediated knockdown and CRISPR/Cas9 gene editing.
Main Results:
- Cep70 is identified as a novel deuterosome-associated protein, forming a complex with CCDC78 and Deup1.
- Cep70 dynamically localizes to deuterosomes during centriole biogenesis.
- Depletion of Cep70 significantly reduces centriole formation and causes defects in MCC radial intercalation.
Conclusions:
- Cep70 is a crucial regulator of deuterosome-mediated centriole biogenesis in MCCs.
- Cep70's function is essential for proper MCC development and tissue organization.
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