PCAF regulates H3 phosphorylation and promotes autophagy in osteosarcoma cells.

Abstract

BACKGROUND:

Osteosarcoma is one of malignant cancer. Histone phosphorylation is common in tumors. We explored the effects of p300-CBP-associated factor (PCAF) and phosphorylation of H3S28 in osteosarcoma cancer cell autophagy.

METHODS:

Osteosarcoma cancer cell lines were collected and/or transfected with full length PCAF or interference miRNAs to mimic or silence of PCAF expression. Immunoprecipitation assay and GST pull down was used to target targeting PCAF or H3S28ph. H3-/- SNU-C1 cells were transfected with H3WT- or H3S28F-expressing or enhanced green fluorescent protein (EGFP)-tagged LC3 plasmids, in which H3 was tagged with HA. An in vitro kinase activity assay was performed to test whether recombinant full-length PCAF could phosphorylate H3 in the site of S28. The functions on autophagy was detected by number of autophagosomes, number of EGFP-LC3, LC3-II/I, percentage of degradation and expression of autophagy associated gene (ATG).

RESULTS:

PCAF positively regulated H3S28ph in osteosarcoma cancer cells; Immunoprecipitation assay and GST pull down demonstrated that PCAF could interact directly with H3 in osteosarcoma cancer cells. In addition, silence of PCAF inhibited the number of autophagosomes, number of EGFP-LC3, LC3-II/I, percentage of degradation and expression of ATG. Moreover, H3S28A (H3S28 mutation) impaired the promoting autophagy effects of PCAF. The PCAF-H3S28ph axis promoted osteosarcoma cancer autophagy viatranscriptional regulation of ATG genes.

CONCLUSION:

PCAF regulated H3S28 phosphorylation and their axis promotes autophagy in osteosarcoma cancer cells viatargeting ATG5 and ATG7.

• Authors: Daliang Kong, Boda Ying, Jinrui Zhang, Hongliang Ying
• Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie (Biomed Pharmacother) Vol. 118 Pg. 109395 (Oct 2019) ISSN: 1950-6007 [Electronic] France
• PMID: 31545241 (Publication Type: Journal Article)
• Copyright: Copyright © 2019. Published by Elsevier Masson SAS.

Chemical References

• Histones
• Phosphoserine
• p300-CBP Transcription Factors
• p300-CBP-associated factor

Topics

• Autophagy
• Cell Line, Tumor
• Cell Proliferation
• Gene Expression Regulation, Neoplastic
• Gene Silencing
• Histones (metabolism)
• Humans
• Osteosarcoma (metabolism, pathology)
• Phosphorylation
• Phosphoserine (metabolism)
• Transcription, Genetic
• p300-CBP Transcription Factors (metabolism)