Abstract
CENP-A is a centromere-specific histone H3 variant that epigenetically determines centromere identity, but how CENP-A is deposited at the centromere remains obscure. We previously reported that CENP-A K124 ubiquitylation, mediated by the CUL4A-RBX1-COPS8 complex, is essential for CENP-A deposition at the centromere. However, a recent report stated that CENP-A K124R mutants show no defects in centromere localization and cell viability. In the present study, we found that EYFP tagging induces additional ubiquitylation of EYFP-CENP-A K124R, which allows the mutant protein to bind to HJURP. Using a previously developed conditional CENP-A knockout system and our CENP-A K124R knockin mutant created by the CRISPR-Cas9 system, we show that the Flag-tagged or untagged CENP-A K124R mutant is lethal. This lethality is rescued by monoubiquitin fusion, indicating that CENP-A ubiquitylation is essential for viability.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 683-689.e6 |
| Journal | Developmental Cell |
| Volume | 50 |
| Issue number | 6 |
| DOIs | |
| State | Published - Sep 23 2019 |
Keywords
- CENP-A
- centromere
- centromere identity
- conditional knockout system
- epigenetics
- kinetochore
- mitosis
- monoubiquitin
- posttranslational modifications (PTMs)
- ubiquitylation
ASJC Scopus subject areas
- Molecular Biology
- Biochemistry, Genetics and Molecular Biology(all)
- Developmental Biology
- Cell Biology