Mobilization and recruitment of HP1: a bimodal response to DNA breakage.
The pathways that signal double-strand DNA breaks (DSBs) in mammalian cells are central to the maintenance of genome integrity. We have reported (Ayoub et al., Nature 2008; 453: 682-6) that the rapid mobilization of the heterochromatin protein, HP1beta, within seconds from DSB sites promotes chromatin changes like H2AX phosphorylation that trigger this response. Notably, this paper and a subsequent report (Ayoub et al., Cell Cycle 2009; 8: 1494-500), demonstrate that transient HP1beta mobilization is followed by its accumulation over time at DSB sites. Indeed, two recent papers (Luijsterburg et al., J Cell Biol 2009; 185:577-86 and Zarebski et al., Cytometry A May 2009) suggest that HP1 recruitment to damage sites, rather than its rapid mobilization, is the predominant behaviour exhibited by this protein. Here, we present new experimental analyses which corroborate that fluorophore-tagged HP1beta exhibits two distinct behaviours at DSB sites in living cells - rapid, transient mobilization, most evident in heterochromatic regions, followed by slower recruitment. Experimental methods allowing visualization of these behaviours are described. Interestingly, chemical inhibition of the DNA-damage responsive enzyme, casein kinase 2 (CK2), suppresses HP1beta mobilization while permitting recruitment. Our findings reconcile recent findings in a new model, wherein rapid HP1beta mobilization from DSBs mediated by its phosphorylation on Thr51 by CK2, is followed by, and may overlap with, its accumulation at these sites via the chromoshadow domain, independent of Thr51. Our analyses provide fresh insight into the earliest events that trigger the DNA damage response in mammalian cells.
Ayoub
Nabieh
N
The Medical Research Council Cancer Cell Unit, Cambridge, UK.
Jeyasekharan
Anand D
AD
Venkitaraman
Ashok R
AR
D000818:Animals; D047390:Casein Kinase II; D000090266:Chromobox Protein Homolog 5; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism* / Q000502:physiology; D053903:DNA Breaks, Double-Stranded; D006801:Humans; D008322:Mammals; D010766:Phosphorylation
C000715593:CBX1 protein, human; D002868:Chromosomal Proteins, Non-Histone; D000090266:Chromobox Protein Homolog 5; D047390:Casein Kinase II