Limits...
A potential role for protein palmitoylation and zDHHC16 in DNA damage response.

Cao N, Li JK, Rao YQ, Liu H, Wu J, Li B, Zhao P, Zeng L, Li J - BMC Mol. Biol. (2016)

Bottom Line: Inhibition of protein palmitoylation compromised DNA damage-induced activation of Atm, induction and activation of p53, cell cycle arrest at G2/M phase, and DNA damage foci assembly/disassembly in primary mouse embryonic fibroblasts.Furthermore, knockout of zDHHC16, a palmitoyltransferase gene identified as an interacting protein for c-Abl, a non-receptor tyrosine kinase involved in DNA damage response, reproduced most of the defects in DNA damage responses produced by the inhibition of protein palmitoylation.Our results revealed critical roles for protein palmitoylation and palmitoyltransferase zDHHC16 in early stages of DNA damage responses and in the regulation of Atm activation.

View Article: PubMed Central - PubMed

Affiliation: Bio-X Institutes, Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Ministry of Education, Shanghai Jiao Tong University, Shanghai, 200240, China.

ABSTRACT

Background: Cells respond to DNA damage by activating the phosphatidylinositol-3 kinase-related kinases, p53 and other pathways to promote cell cycle arrest, apoptosis, and/or DNA repair. Here we report that protein palmitoylation, a modification carried out by protein acyltransferases with zinc-finger and Asp-His-His-Cys domains (zDHHC), is required for proper DNA damage responses.

Results: Inhibition of protein palmitoylation compromised DNA damage-induced activation of Atm, induction and activation of p53, cell cycle arrest at G2/M phase, and DNA damage foci assembly/disassembly in primary mouse embryonic fibroblasts. Furthermore, knockout of zDHHC16, a palmitoyltransferase gene identified as an interacting protein for c-Abl, a non-receptor tyrosine kinase involved in DNA damage response, reproduced most of the defects in DNA damage responses produced by the inhibition of protein palmitoylation.

Conclusions: Our results revealed critical roles for protein palmitoylation and palmitoyltransferase zDHHC16 in early stages of DNA damage responses and in the regulation of Atm activation.

No MeSH data available.


Related in: MedlinePlus

Inhibition of palmitoylation impaired DNA damage-induced cell cycle arrest at G2/M phase. Primary MEFs were pre-treated with 50 μM 2BP for 24 h and then treated with 1 μM of Dox for 24 h. The cells were harvested, fixed, stained with PI, and analyzed by flow cytometry. a Representative micrographs of flow cytometry analysis. b Averaged data from multiple experiments. The total number of cells analyzed was set as 100 % and cells at different phase were calculated accordingly
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4862184&req=5

Fig5: Inhibition of palmitoylation impaired DNA damage-induced cell cycle arrest at G2/M phase. Primary MEFs were pre-treated with 50 μM 2BP for 24 h and then treated with 1 μM of Dox for 24 h. The cells were harvested, fixed, stained with PI, and analyzed by flow cytometry. a Representative micrographs of flow cytometry analysis. b Averaged data from multiple experiments. The total number of cells analyzed was set as 100 % and cells at different phase were calculated accordingly

Mentions: DNA damage eventually induces apoptosis or cell cycle arrest. Since 2BP compromised Atm activation and p53 induction and activation, we suspected that 2BP might affect Dox-induced cell cycle arrest and apoptosis. Cell cycle arrest was analyzed by flow cytometry (Fig. 5a). Dox treatment led to an accumulation of cells in the G2/M phase, a decrease in G1 phase cells, and a modest decrease in S phase cells in wild type MEFs. 2BP pre-treatment impeded Dox-induced increase in the percentage of cells arrested in the G2/M phase compared to control cells, without altering the percentage of S phase cells (Fig. 5b). These results suggest that protein palmitoylation is required for proper G2/M cell cycle arrest in DNA damage response. However, we found that 2BP showed some toxicity to MEFs at the doses used to inhibit protein palmitoylation, which may reflect a requirement for protein palmitoylation for cell survival, and the combination of 2BP and Dox caused more cell death (Additional file 1: Figure S3). This prevented us from further testing the role of 2BP-suppressed p53 activation in DNA damage-induced apoptosis.Fig. 5


A potential role for protein palmitoylation and zDHHC16 in DNA damage response.

Cao N, Li JK, Rao YQ, Liu H, Wu J, Li B, Zhao P, Zeng L, Li J - BMC Mol. Biol. (2016)

Inhibition of palmitoylation impaired DNA damage-induced cell cycle arrest at G2/M phase. Primary MEFs were pre-treated with 50 μM 2BP for 24 h and then treated with 1 μM of Dox for 24 h. The cells were harvested, fixed, stained with PI, and analyzed by flow cytometry. a Representative micrographs of flow cytometry analysis. b Averaged data from multiple experiments. The total number of cells analyzed was set as 100 % and cells at different phase were calculated accordingly
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4862184&req=5

Fig5: Inhibition of palmitoylation impaired DNA damage-induced cell cycle arrest at G2/M phase. Primary MEFs were pre-treated with 50 μM 2BP for 24 h and then treated with 1 μM of Dox for 24 h. The cells were harvested, fixed, stained with PI, and analyzed by flow cytometry. a Representative micrographs of flow cytometry analysis. b Averaged data from multiple experiments. The total number of cells analyzed was set as 100 % and cells at different phase were calculated accordingly
Mentions: DNA damage eventually induces apoptosis or cell cycle arrest. Since 2BP compromised Atm activation and p53 induction and activation, we suspected that 2BP might affect Dox-induced cell cycle arrest and apoptosis. Cell cycle arrest was analyzed by flow cytometry (Fig. 5a). Dox treatment led to an accumulation of cells in the G2/M phase, a decrease in G1 phase cells, and a modest decrease in S phase cells in wild type MEFs. 2BP pre-treatment impeded Dox-induced increase in the percentage of cells arrested in the G2/M phase compared to control cells, without altering the percentage of S phase cells (Fig. 5b). These results suggest that protein palmitoylation is required for proper G2/M cell cycle arrest in DNA damage response. However, we found that 2BP showed some toxicity to MEFs at the doses used to inhibit protein palmitoylation, which may reflect a requirement for protein palmitoylation for cell survival, and the combination of 2BP and Dox caused more cell death (Additional file 1: Figure S3). This prevented us from further testing the role of 2BP-suppressed p53 activation in DNA damage-induced apoptosis.Fig. 5

Bottom Line: Inhibition of protein palmitoylation compromised DNA damage-induced activation of Atm, induction and activation of p53, cell cycle arrest at G2/M phase, and DNA damage foci assembly/disassembly in primary mouse embryonic fibroblasts.Furthermore, knockout of zDHHC16, a palmitoyltransferase gene identified as an interacting protein for c-Abl, a non-receptor tyrosine kinase involved in DNA damage response, reproduced most of the defects in DNA damage responses produced by the inhibition of protein palmitoylation.Our results revealed critical roles for protein palmitoylation and palmitoyltransferase zDHHC16 in early stages of DNA damage responses and in the regulation of Atm activation.

View Article: PubMed Central - PubMed

Affiliation: Bio-X Institutes, Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Ministry of Education, Shanghai Jiao Tong University, Shanghai, 200240, China.

ABSTRACT

Background: Cells respond to DNA damage by activating the phosphatidylinositol-3 kinase-related kinases, p53 and other pathways to promote cell cycle arrest, apoptosis, and/or DNA repair. Here we report that protein palmitoylation, a modification carried out by protein acyltransferases with zinc-finger and Asp-His-His-Cys domains (zDHHC), is required for proper DNA damage responses.

Results: Inhibition of protein palmitoylation compromised DNA damage-induced activation of Atm, induction and activation of p53, cell cycle arrest at G2/M phase, and DNA damage foci assembly/disassembly in primary mouse embryonic fibroblasts. Furthermore, knockout of zDHHC16, a palmitoyltransferase gene identified as an interacting protein for c-Abl, a non-receptor tyrosine kinase involved in DNA damage response, reproduced most of the defects in DNA damage responses produced by the inhibition of protein palmitoylation.

Conclusions: Our results revealed critical roles for protein palmitoylation and palmitoyltransferase zDHHC16 in early stages of DNA damage responses and in the regulation of Atm activation.

No MeSH data available.


Related in: MedlinePlus