Chromosome missegregation during anaphase triggers p53 cell cycle arrest through histone H3.3 Ser31 phosphorylation

Abstract

Maloriented chromosomes can evade the spindle assembly checkpoint and generate aneuploidy, a common feature of tumorigenesis. But chromosome missegregation in non-transformed cells triggers a p53-dependent fail-safe mechanism that blocks proliferation of normal cells that inadvertently become aneuploid. How this fail-safe is triggered is not known. Here we identify a conserved feedback mechanism that monitors missegregating chromosomes during anaphase through the differential phosphorylation of histone H3.3 at Ser31. We do this by inducing transient chromosome missegregation in diploid cells. During anaphase, H3.3 Ser31 is phosphorylated along the arms of lagging or misaligned chromosomes. Within minutes, Ser31 phosphorylation (Ser31P) spreads to all of the chromatids of both daughter cells, which persists into G1. Masking H3.3 Ser31P by antibody microinjection prevents nuclear p53 accumulation in the aneuploid daughters. Previous work demonstrated that prolonged prometaphase and DNA damage during abnormal mitosis can activate p53. We show that p53 activation in response to chromosome missegregation can occur without prolonged mitosis or DNA damage. Our study provides insight into how aneuploidy caused by chromosome missegregation is normally monitored and suppressed.

BCL9L Dysfunction Impairs Caspase-2 Expression Permitting Aneuploidy Tolerance in Colorectal Cancer

“…Second, we performed a screen to detect tolerance of chromosome segregation errors. Chromosome missegregation induces p53-mediated cell cycle arrest in the next G1 phase, often followed by apoptosis, thereby preventing propagation of aneuploid progeny (Figure S3A; see also Hinchcliffe et al., 2016). Chromosome missegregation can be artificially induced in HCT-116 cells with reversine, an Mps1 inhibitor that impairs the spindle assembly checkpoint resulting in chromosomal non-disjunction and missegregation.…”

“…DNA damage (Janssen et al., 2011), histone phosphorylation (Hinchcliffe et al., 2016) and reactive oxygen species (Li et al., 2010) have all been proposed as mechanisms of p53 accumulation in CIN cells. Our data reveal that caspase-2 depletion induces tolerance of endogenous chromosome segregation errors and prevents p53 accumulation in response to artificial induction of chromosome segregation errors using an Mps1 inhibitor, reversine, supporting a central role for caspase-2 as an enzyme regulating p53, underpinned by seminal work from other groups (Dorstyn et al., 2012, Oliver et al., 2011, Terry et al., 2015).…”

The PIDDosome activates p53 in response to supernumerary centrosomes

“…p53 appears the most relevant effector responding to mitotic defects, inducing cell cycle arrest or cell death following aberrant mitoses (Vitale et al 2011). The triggers of mitotic defects can be very heterogeneous and include (1) DNA damage, occurring as either a consequence of sublethal caspase activation on extended mitosis (Orth et al 2012) or a result of chromosome segregation defects (Janssen et al 2011;Crasta et al 2012); (2) the extension of the mitotic duration itself above a critical threshold (Uetake and Sluder 2010;Fong et al 2016;Lambrus et al 2016;Meitinger et al 2016); (3) chromosome congression/segregation defects (Thompson and Compton 2010;Hinchcliffe et al 2016); and (4) cytokinesis failure or centrosome amplification (Holland et al 2012;Ganem et al 2014). While the first three cues appear clearly distinct from each other, either requiring a definite set of genetic factors for p53 activation or associating with specific markers, it remained elusive whether the presence of extra centrosomes suffices to trigger p53 activation or whether this occurs as a consequence of the resulting CIN.…”

Living in CIN: Mitotic Infidelity and Its Consequences for Tumor Promotion and Suppression

“…Similarly, tetraploidy was reported to stabilize p53 through oxidative DNA damage that did not correlate with γH2AX staining, but was dependent on the DNA damage checkpoint kinase ATM (Kuffer et al, 2013). A recent study reported that Serine 31 on histone H3 is phosphorylated on missegregated chromosomes that are separated from the masses of segregating DNA during anaphase, and that this phosphorylation was sufficient to stabilize p53 in the subsequent G1 in a manner independent of the DNA damage checkpoint kinases ATM, ATR and Chk1 (Hinchcliffe et al, 2016). …”