11E

Amino Acid Sequence

3D Structure (/structures/AF-P09874-F1-model_v6.cif)

Function

Poly-ADP-ribosyltransferase that mediates poly-ADP-ribosylation of proteins and plays a key role in DNA repair. Mediates glutamate, aspartate, serine, histidine or tyrosine ADP-ribosylation of proteins: the ADP-D-ribosyl group of NAD+ is transferred to the acceptor carboxyl group of target residues and further ADP-ribosyl groups are transferred to the 2'-position of the terminal adenosine moiety, building up a polymer with an average chain length of 20-30 units. Serine ADP-ribosylation of proteins constitutes the primary form of ADP-ribosylation of proteins in response to DNA damage. Specificity for the different amino acids is conferred by interacting factors, such as HPF1 and NMNAT1. Following interaction with HPF1, catalyzes serine ADP-ribosylation of target proteins; HPF1 confers serine specificity by completing the PARP1 active site. Also catalyzes tyrosine ADP-ribosylation of target proteins following interaction with HPF1. Following interaction with NMNAT1, catalyzes glutamate and aspartate ADP-ribosylation of target proteins; NMNAT1 confers glutamate and aspartate specificity (By similarity). PARP1 initiates the repair of DNA breaks: recognizes and binds DNA breaks within chromatin and recruits HPF1, licensing serine ADP-ribosylation of target proteins, such as histones (H2BS6ADPr and H3S10ADPr), thereby promoting decompaction of chromatin and the recruitment of repair factors leading to the reparation of DNA strand breaks. HPF1 initiates serine ADP-ribosylation but restricts the polymerase activity of PARP1 in order to limit the length of poly-ADP-ribose chains. In addition to base excision repair (BER) pathway, also involved in double-strand breaks (DSBs) repair: together with TIMELESS, accumulates at DNA damage sites and promotes homologous recombination repair by mediating poly-ADP-ribosylation. Mediates the poly-ADP-ribosylation of a number of proteins, including itself, APLF, CHFR, RPA1 and NFAT5. In addition to proteins, also able to ADP-ribosylate DNA: catalyzes ADP-ribosylation of DNA strand break termini containing terminal phosphates and a 2'-OH group in single- and double-stranded DNA, respectively. Required for PARP9 and DTX3L recruitment to DNA damage sites. PARP1-dependent PARP9-DTX3L-mediated ubiquitination promotes the rapid and specific recruitment of 53BP1/TP53BP1, UIMC1/RAP80, and BRCA1 to DNA damage sites. PARP1-mediated DNA repair in neurons plays a role in sleep: senses DNA damage in neurons and promotes sleep, facilitating efficient DNA repair (By similarity). In addition to DNA repair, also involved in other processes, such as transcription regulation, programmed cell death, membrane repair, adipogenesis and innate immunity. Acts as a repressor of transcription: binds to nucleosomes and modulates chromatin structure in a manner similar to histone H1, thereby altering RNA polymerase II. Acts both as a positive and negative regulator of transcription elongation, depending on the context. Acts as a positive regulator of transcription elongation by mediating poly-ADP-ribosylation of NELFE, preventing RNA-binding activity of NELFE and relieving transcription pausing. Acts as a negative regulator of transcription elongation in response to DNA damage by catalyzing poly-ADP-ribosylation of CCNT1, disrupting the phase separation activity of CCNT1 and subsequent activation of CDK9. Involved in replication fork progression following interaction with CARM1: mediates poly-ADP-ribosylation at replication forks, slowing fork progression. Poly-ADP-ribose chains generated by PARP1 also play a role in poly-ADP-ribose-dependent cell death, a process named parthanatos (By similarity). Also acts as a negative regulator of the cGAS-STING pathway. Acts by mediating poly-ADP-ribosylation of CGAS: PARP1 translocates into the cytosol following phosphorylation by PRKDC and catalyzes poly-ADP-ribosylation and inactivation of CGAS. Acts as a negative regulator of adipogenesis: catalyzes poly-ADP-ribosylation of histone H2B on 'Glu-35' (H2BE35ADPr) following interaction with NMNAT1, inhibiting phosphorylation of H2B at 'Ser-36' (H2BS36ph), thereby blocking expression of pro-adipogenetic genes (By similarity). Involved in the synthesis of ATP in the nucleus, together with NMNAT1, PARG and NUDT5. Nuclear ATP generation is required for extensive chromatin remodeling events that are energy-consuming.

Mechanism

The ubiquitin-proteasome pathway (UPP) constitutes the principal pathway for protein degradation in eukaryotic cells, mediating the breakdown of target proteins through ubiquitin tagging and subsequent proteasomal degradation. In an ATP-dependent manner, the ubiquitin-activating enzyme (E1) activates ubiquitin (Ub) and conjugates it to a ubiquitin-conjugating enzyme (E2). E3 ligase subsequently catalyzes the transfer of Ub to the target protein, leading to the formation of a polyubiquitin chain that completes the ubiquitination mark. The 19S regulatory particle of the 26S proteasome recognizes this ubiquitination signal and, after deubiquitination, mediates the ATP-dependent delivery of the substrate into the 20S proteasome for degradation.

Description

To investigate the mechanism of PARP1 degradation mediated by 11e, we employed a proteasome inhibition assay. Results demonstrated that the proteasome inhibitor MG132 suppressed 11e-mediated PARP1 degradation. Conversely, when 11e was co-administered with the lysosomal chemotactic agent chloroquine (CQ), CQ did not interfere with 11e-mediated PARP1 degradation (Fig. 5). These findings revealed that the UPS participated in the 11e-mediated degradation of PARP1.

Description

To evaluate the efficacy of HyT-based PARP1 degraders, we investigated the degradation efficiency of compound 11e on PARP1 in MDA- MB-231 cells. The results demonstrated a concentration-dependent degradation of PARP1 by compound 11e, with a DC₅₀ value of 2.16 ± 0.29 μM (Fig. 4A, B).

Description

Generally compared to compounds 11a–11e, compounds 14a–14e exhibited weaker antiproliferative activity, indicating that carbonyl group might be favorable for molecular interactions. Among them, compound 11e showed potent inhibitory activity in MDA-MB-231 cells (IC₅₀ = 4.18±0.30 μM), which was more than twofold higher than that of the positive control drug Olaparib (IC₅₀ = 10.03±0.07 μM)..

Description

Through comprehensive SAR analysis, compound 11e was identified as the most effective PARP1 degrader, exhibiting potent inhibitory activity in both breast and colon cancer cell lines. Compound 11e exhibited potent antitumor activity, with IC₅₀ values of 4.18 ± 0.30 μM, 11.15 ± 0.18 μM, and 6.96 ± 0.12 μM in MDA-MB-231, SW620, and MDA-MB-468 cells, respectively. Its activity in MDA-MB-231 cells was significantly superior to that of Olaparib (10.03 ± 0.07 μ M).

Description

Through comprehensive SAR analysis, compound 11e was identified as the most effective PARP1 degrader, exhibiting potent inhibitory activity in both breast and colon cancer cell lines. Compound 11e exhibited potent antitumor activity, with IC₅₀ values of 4.18 ± 0.30 μM, 11.15 ± 0.18 μM, and 6.96 ± 0.12 μM in MDA-MB-231, SW620, and MDA-MB-468 cells, respectively. Its activity in MDA-MB-231 cells was significantly superior to that of Olaparib (10.03 ± 0.07 μ M).