HP1α is a chromatin crosslinker that controls nuclear and mitotic chromosome mechanics. Author Amy Strom, Ronald Biggs, Edward Banigan, Xiaotao Wang, Katherine Chiu, Cameron Herman, Jimena Collado, Feng Yue, Joan Politz, Leah Tait, David Scalzo, Agnes Telling, Mark Groudine, Clifford Brangwynne, John Marko, Andrew Stephens Publication Year 2021 Type Journal Article Abstract Chromatin, which consists of DNA and associated proteins, contains genetic information and is a mechanical component of the nucleus. Heterochromatic histone methylation controls nucleus and chromosome stiffness, but the contribution of heterochromatin protein HP1α (CBX5) is unknown. We used a novel HP1α auxin-inducible degron human cell line to rapidly degrade HP1α. Degradation did not alter transcription, local chromatin compaction, or histone methylation, but did decrease chromatin stiffness. Single-nucleus micromanipulation reveals that HP1α is essential to chromatin-based mechanics and maintains nuclear morphology, separate from histone methylation. Further experiments with dimerization-deficient HP1α indicate that chromatin crosslinking via HP1α dimerization is critical, while polymer simulations demonstrate the importance of chromatin-chromatin crosslinkers in mechanics. In mitotic chromosomes, HP1α similarly bolsters stiffness while aiding in mitotic alignment and faithful segregation. HP1α is therefore a critical chromatin-crosslinking protein that provides mechanical strength to chromosomes and the nucleus throughout the cell cycle and supports cellular functions. Keywords Humans, Cell Line, Cell Nucleus, Chromosomal Proteins, Non-Histone, Mitosis, Methylation, Chromatin, Chromosomes, Chromobox Protein Homolog 5 Journal Elife Volume 10 Date Published 2021 Jun 09 ISSN Number 2050-084X DOI 10.7554/eLife.63972 Alternate Journal Elife PMCID PMC8233041 PMID 34106828 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML