ATM induces MacroD2 nuclear export upon DNA damage

Abstract

ADP-ribosylation is a dynamic post-translation modification that regulates the early phase of various DNA repair pathways by recruiting repair factors to chromatin. ADP-ribosylation levels are defined by the activities of specific transferases and hydrolases. However, except for the transferase PARP1/ARDT1 little is known about regulation of these enzymes. We found that MacroD2, a mono-ADP-ribosylhydrolase, is exported from the nucleus upon DNA damage, and that this nuclear export is induced by ATM activity. We show that the export is dependent on the phosphorylation of two SQ/TQ motifs, suggesting a novel direct interaction between ATM and ADP-ribosylation. Lastly, we show that MacroD2 nuclear export temporally restricts its recruitment to DNA lesions, which may decrease the net ADP-ribosylhydrolase activity at the site of DNA damage. Together, our results identify a novel feedback regulation between two crucial DNA damage-induced signaling pathways: ADP-ribosylation and ATM activation.

(ADP-ribosyl)hydrolases: structure, function, and biology

“…Furthermore, other studies suggest that they can also hydrolyze OAADPr as well as (ADP-ribosyl) ated nucleic acids Munnur and Ahel 2017;Agnew et al 2018;Munnur et al 2019). MacroD1 localizes largely to the mitochondrial matrix (Agnew et al 2018), whereas MacroD2 distributes in the cytosol and nucleus Golia et al 2017). The physiological substrates and cellular functions of both MacroD1 and MacroD2 remain largely elusive.…”

“…Alterations of MacroD2 functions in the DNA damage response and signal transduction may also be linked to tumor formation and/or progression. Indeed, MacroD2 is phosphorylated by ATM in response to DNA doublestrand breaks, as well as being involved in reversing the ADP-ribosylation of GSK3β, a key kinase involved in the WNT-mediated signal transduction pathway Golia et al 2017).…”

ADP‐ribosylation: new facets of an ancient modification

“…Some macrodomains have also evolved enzymatic activity and are capable of hydrolysing ADP‐ribosylation (see below) . As a consequence, macrodomain‐containing proteins are involved in a diverse set of cellular functions, such as chromatin remodelling and DNA‐damage repair, oxidative stress response, metabolic processes and pathogenic mechanisms . In addition to macrodomain, several other widely distributed domains have been described as readers for ADP‐ribosylation, such as the PAR‐binding zinc finger (PBZ) , the WWE (named after three of its conserved residues) , the oligonucleotide/oligosaccharide‐binding (OB) domain and the PAR‐binding motifs (PBM) which is abundant in DNA‐damage repair proteins .…”

The Controversial Roles of ADP-Ribosyl Hydrolases MACROD1, MACROD2 and TARG1 in Carcinogenesis

“…However, it was not tested whether the identified compound might inhibit one of the macrodomains with catalytic activity [69]. [40,70,71] Ubiquitous expression, enriched in skeletal muscle [70,71] mRNA expression is induced by oestrogen [72][73][74] MACROD2 C20orf133 MACROD2 47 kDa 50 kDa [70] Diffuse nuclear, cytoplasmic [70,75] So far detected only in the brain [70,76] Phosphorylation by ATM upon DNA damage induces translocation to cytoplasm [75] TARG1 C6orf130 OARD1 17 kDa 17 kDa [14,70] Nuclear, nucleolar, stress granular [14,45,70] Ubiquitous expression [70] Leaves nucleoli upon DNA damage [14] * The predicted molecular weight of MACROD1 is 35 kDa. However, due to cleavage of the N-terminus upon translocation into mitochondria, the protein detected in western blotting (WB) is smaller [70].…”

“…MACROD1 resides in mitochondria [40,70,71], but, depending on the tag used for overexpression, will reside exclusively in the nucleus instead [70]. MACROD2 displays a more diffuse cytoplasmic/nuclear distribution [70,75] and overexpressed TARG1 appears to be present in nucleoplasm, nucleoli, and cytoplasmic stress granules [14,70]. TARG1 and MACROD1 are expressed throughout the different tissues with an enrichment of MACROD1 in skeletal muscle at both the protein and RNA level [70].…”