Long-term ethanol consumption can contribute to the development of many diseases, including cardiovascular and cancer. intermediate is proposed to be carried out by a water molecule that is activated by the His493 residue acting as a general base. It was known that phospholipase D enzymes are able to catalyze not only hydrolysis but also a transphosphatidylation reaction in the presence of primary alcohols in which they transfer the substrate to the alcohol instead of water. Here, we first exhibited that TDP1 is able to undergo a transphosphooligonucleotidation reaction, transferring the substrate residue to the alcohol, thus inducing the formation of covalent DNA adducts with different primary alcohol residues. Such adducts can be accumulated in the conditions of high concentration of alcohol. We exhibited that glycerol residue was efficiently cleaved from the 3-end by TDP1 but not by its mutant Rabbit Polyclonal to SFRS4 form associated with the disease spinocerebellar ataxia with axonal neuropathy. Therefore, the second reaction step can be carried out not only by a water molecule but also by the other small nucleophilic molecules, e.g., glycerol and ethanol. Thus, in some cases, TDP1 can be regarded not only as a repair enzyme but also as a source of DNA damage especially in the case of mutation. Such damages can make a negative contribution to the stability of cell vitality. as repairing the covalently linked adducts of DNA topoisomerase I (TOP1) by catalyzing the hydrolysis of the phosphodiester bond between the tyrosine residue of TOP1 peptide and the 3 phosphate of DNA. The result DNA product has a break with 3 phosphate and 5 hydroxyl groups (Yang et al., 1996; Pouliot et al., 1999). TDP1 possesses a unique HKD motif that differs from other PLD superfamily members, and its orthologs represent a distinct class within the PLD superfamily. TDP1 catalytic center contains two histidine residues His493 and His263 (Interthal et al., 2001). The His493Arg mutation in Tdp1 gene causes spinocerebellar ataxia with axonal neuropathy type 1 (SCAN1) by affecting neuronal cells (Takashima et al., 2002). TDP1 activity is not limited by the removal of cellular TOP1 adducts. TDP1 was shown to catalyze 3 phosphoglycolate removal from a single-stranded oligonucleotide and a single strand overhangs of DNA double-strand breaks (Inamdar et al., 2002; Raymond et al., 2005). TDP1 is now regarded as a general 3 DNA end-processing enzyme that acts within the single-strand break repair complex to remove adducts and to prepare the DNA ends bearing 3 phosphate group for further processing by DNA repair enzymes (Rass et al., 2007). TDP1 also possesses a DNA and RNA 3-nucleosidase activity that removes from the Sesamin (Fagarol) 3-end of the substrate a single nucleoside, as well as nucleoside analogs terminating DNA synthesis and widely used as antiviral and anticancer brokers and a variety of synthetic DNA adducts for example with molecules, such as biotin and various fluorophores (Dexheimer et al., 2008; Murai et al., 2012; Huang et al., 2013; Dyrkheeva et al., 2018; Brettrager and van Waardenburg, 2019). TDP1 can also Sesamin (Fagarol) process other 3 DNA end blocking lesions as a substrate: 3 abasic sites (tetrahydrofuran and ,-unsaturated aldehyde) and different bulky substituents (Hawkins et al., 2009; Interthal et al., 2005a; Zhou et al., 2005). TDP1 can reverse not only 3-TOP1-DNA cross-linked bonds but also it is able to release different DNA-protein cross-links. It was found that both Sesamin (Fagarol) human and yeast TDP1 proteins have the ability to process 5-phosphotyrosyl and 5-phosphotyrosyl-linked peptide substrates, thus indicating that they can hydrolyze covalently linked adducts of DNA with TOP2 (Nitiss et al., 2006; Murai et al., 2012; Zhang et al., 2020). It also works on the other large adducts including protein fragments (peptides) as a result of failed Schiff base linked proteins, such as Sesamin (Fagarol) proteolytically processed poly(ADP-ribose) polymerase 1 Sesamin (Fagarol) (PARP1)-DNA adducts. These different protein-DNA adducts can be stabilized by chemotherapeutic compounds, e.g., camptothecins, etoposide, and local DNA perturbations introduced by irradiation and endogenous reactive oxygen species (Brettrager and van Waardenburg, 2019). We have previously shown that human TDP1 can also cleave an apurinic/apyrimidinic (AP) site and its synthetic analogs located inside DNA strand with the formation of 3 phosphate termini. This observation allows suggesting a novel pathway of AP site repair impartial of AP endonuclease 1 (APE1) (Lebedeva et al., 2011, 2012, 2013; Kuznetsov et al., 2017). In contrast to APE1, TDP1 more effectively hydrolyzes AP sites in single-stranded DNA than in DNA duplex (Lebedeva et al., 2012). This suggests that TDP1 may be.