Stability of centrosome numbers in poly(ADP-ribose) polymerase-1- and poly(ADP-ribose) glycohydrolase-deficient mouse ES cells

Abstract

Poly(ADP-ribose) polymerase-1 (Parp-1) localizes mainly in the nucleus and functions in DNA repair, genome stability and cell death regulation. Meanwhile, it also localizes in centrosomes and is involved in the regulation of centrosome duplication. An abnormal increase in centrosome numbers is frequently observed in Parp-1-deficient (Parp-1-/-) mouse embryonic fibroblasts (MEFs) (Kanai et al. (2003) Mol. Cell. Biol. 23, 2451-2462). However, there are no studies on whether the centrosome abnormality occurs also in other cell types under Parp-1 deficiency. In this study, we report that Parp-1-/- mouse embryonic stem (ES) cell lines did not show an abnormally increased number of centrosomes compared to wild-type ES cells. Recently, poly(ADP-ribose) glycohydrolase (Parg) has also been shown to localize in centrosomes (Ohashi et al. (2003) Biochem. Biophys. Res. Commun. 307, 915-921). The number of centrosomes of Parg-deficient (Parg-/-) ES cells was also analyzed in this study and was found to be stable under Parg deficiency. We also examined centrosome numbers in wild-type, Parp-1-/- and Parg-/- ES cell lines after treatment with methylmethanesulfonate (MMS) or γ-irradiation. Although a slight increase in the number of centrosomes is observed in each genotype twenty-four hours after treatment with MMS at 50 μM or with γ-irradiation at 1.4 Gy, there was no difference among the genotypes. These results suggest that loss of Parp-1 and Parg is insufficient to induce abnormality in centrosome numbers in ES cells and that ES cells possibly possess a strict mechanism for the maintenance of a normal number of centrosomes. (Contributed by Takashi SUGIMURA, M.J.A.)