Orderly mitosis shapes interphase genome architecture

Area of Science:

• Cell Biology
• Genomics
• Molecular Biology

Background:

• The 3D genome architecture within the interphase nucleus is complex.
• The molecular mechanisms governing global genome organization remain largely unknown.

Purpose of the Study:

• To identify molecular mechanisms driving higher-order genome organization.
• To understand the regulation of spatial genome architecture.

Main Methods:

• Conducted high-throughput imaging-based CRISPR knockout screens targeting 1064 nuclear protein-coding genes.
• Utilized centromere distribution at single-cell resolution as a surrogate marker for genome organization.
• Employed multiple human cell lines for comprehensive analysis.

Main Results:

• Identified major regulators of spatial centromere distribution, including nucleolar, kinetochore, cohesin, condensin, and nuclear pore complex components.
• Demonstrated that centromere distribution changes require cell cycle progression.
• Showed that depletion of specific mitotic factors alters centromere distribution in the subsequent interphase.

Conclusions:

• Identified key molecular determinants of spatial centromere organization.
• Established that ordered progression through mitosis is crucial for shaping interphase genome architecture.