J 2023

Irradiation potentiates p53 phosphorylation and p53 binding to the promoter and coding region of the TP53 gene

LEGARTOVÁ, Soňa, Paolo FAGHERAZZI, Pratik GOSWAMI, Václav BRÁZDA, Gabriela LOCHMANOVÁ et. al.

Základní údaje

Originální název

Irradiation potentiates p53 phosphorylation and p53 binding to the promoter and coding region of the TP53 gene

Autoři

LEGARTOVÁ, Soňa, Paolo FAGHERAZZI (380 Itálie, domácí), Pratik GOSWAMI (356 Indie, domácí), Václav BRÁZDA, Gabriela LOCHMANOVÁ (203 Česká republika, domácí), Irena KOUTNÁ a Eva BÁRTOVÁ (garant)

Vydání

Biochimie, Elsevier, 2023, 0300-9084

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Francie

Utajení

není předmětem státního či obchodního tajemství

Odkazy

Impakt faktor

Impact factor: 3.900 v roce 2022

Kód RIV

RIV/00216224:14740/23:00130443

Organizační jednotka

Středoevropský technologický institut

UT WoS

000921740700001

Klíčová slova anglicky

p53; 53BP1; DNA damage; Epigenetics; Mass spectrometry; FLIM-FRET; AFM

Štítky

Příznaky

Mezinárodní význam, Recenzováno
Změněno: 9. 10. 2024 13:35, Ing. Martina Blahová

Anotace

V originále

An essential factor of the DNA damage response is 53BP1, a multimeric protein that inhibits the resection-dependent double-strand break (DBS) repair. The p53 protein is a tumor suppressor known as a guardian of the genome. Although the interaction between 53BP1 and its p53 partner is well-known in regulating gene expression, a question remains whether genome injury can affect the interaction between 53BP1 and p53 proteins or p53 binding to DNA. Here, using mass spectrometry, we determine post-translational modifications and interaction properties of 53BP1 and p53 proteins in non-irradiated and γ-irradiated cells. In addition, we used Atomic Force Microscopy (AFM) and Fluorescent Lifetime Imaging Microscopy combined with Fluorescence Resonance Energy Transfer (FLIM-FRET) for studies of p53 binding to DNA. Also, we used local laser microirradiation as a tool of advanced confocal microscopy, showing selected protein accumulation at locally induced DNA lesions. We observed that 53BP1 and p53 proteins accumulate at microirradiated chromatin but with distinct kinetics. The density of 53BP1 (53BP1pS1778) phosphorylated form was lower in DNA lesions than in the non-specified form. By mass spectrometry, we found 22 phosphorylations, 4 acetylation sites, and methylation of arginine 1355 within the DNA-binding domain of the 53BP1 protein (aa1219-1711). The p53 protein was phosphorylated on 8 amino acids and acetylated on the N-terminal domain. Post-translational modifications (PTMs) of 53BP1 were not changed in cells exposed to γ-radiation, while γ-rays increased the level of S6ph and S15ph in p53. Interaction analysis showed that 53BP1 and p53 proteins have 54 identical interaction protein partners, and AFM revealed that p53 binds to both non-specific and TP53-specific sequences (AGACATGCCTA GGCATGTCT). Irradiation by γ-rays enhanced the density of the p53 protein at the AGACATGCCTAGGCATGTCT region, and the binding of p53 S15ph to the TP53 promoter was potentiated in irradiated cells. These findings show that γ-irradiation, in general, strengthens the binding of phosphorylated p53 protein to the encoding gene.

Návaznosti

EF18_046/0015974, projekt VaV
Název: Modernizace České infrastruktury pro integrativní strukturní biologii
GF19-29701L, projekt VaV
Název: Funkce HDAC1 v T-buněčných lymfomech
Investor: Grantová agentura ČR, Funkce HDAC1 v T-buněčných lymfomech, Partnerská agentura (Rakousko)
90127, velká výzkumná infrastruktura
Název: CIISB II