Target Proteins

POP1

POP1 (Processing of Precursors-1) is a ribonuclease that is most well-known for its vital role in the RNase P and MRP complexes. These highly conserved complexes have vital roles in the cell, as RNase P matures the 5’ ends of tRNA and RNase MRP cleaves pre-rRNA (1,2). In both of these, POP1 binds simultaneously with POP6/7 to the RNA components of the complex. This provides protection to specific parts of the RNA, encouraging proteins like RNase A and RNase VI to cleave the unprotected regions (3).

Recently, POP1 has been implicated in the formation of the telomerase complex. It is suggested to use its RNA binding ability to stabilise TERC (Telomerase RNA Component), in order for the telomerase complex to be recruited to the RNA (4). This has been proposed via the P3 domain of the RNA, which is significantly homologous between TLC1, the specific RNA strand of TERC that POP1 would bind to, NME1, the RNA portion of RNase MRP, and RPR1, the RNA component of RNase P. In fact, these P3 regions have been said to be functionally interchangeable in yeast (5). These regions are highly conserved throughout eukaryotes, so it is thought that this discovery reflects the human structures of these proteins. In fact, POP1 upregulation has been found as a factor in breast cancer development, theoretically because of the upregulation of telomerase and extension of the telomeres of the cancer cell (4). This makes it a great target for our PROTAC design.

POP1’s Saccharomyces cerevisiae homolog of the same name shows very close structural and functional homology as the human protein. The two homologs have also been shown to be evolutionarily related (6).

ATM

Ataxia-Talengiectasia Mutated (ATM) is a serine/thronine pleiotropic protein kinase that plays a key role in survival (7). Activation of this protein is involved in cell cycle checkpoint signalling, double strand break (DSB) repair, apoptosis initiation, and genome surveillance (8,9). Most importantly, ATM mediates the preferential recruitment of the telomerase complex to shorter telomeres (812).

ATM is part of the phosphatidylinositol 3-kinase-related kinase (PIKK) family of serine/threonine protein kinases (13). Members of this family share similar structural and functional properties, and each have conserved sequence homology in their C-terminal kinase domain, FAT domain, and and FATC domain (7,14).

ATM’s close yeast homolog, Tel1, has been shown to be responsible for the preferential recuruitment of the telomerase complex to shorter telomeres in Saccharomyces cerevisiae (7,1517). Fritz and colleagues (2000) showed that human cells derived from ataxia-talengiectasia patients showed relieved symptoms in response to overexpression of Tel1, the yeast homologue (18). This demonstrates their structural and functional homology.

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References

1.
Wu J, Niu S, Tan M, Huang C, Li M, Song Y, et al. Cryo-EM structure of the human ribonuclease p holoenzyme. Cell [Internet]. 2018;175(5):1393–1404.e11. Available from: https://www.sciencedirect.com/science/article/pii/S0092867418313114
2.
3.
Fagerlund RD, Perederina A, Berezin I, Krasilnikov AS. Footprinting analysis of interactions between the largest eukaryotic RNase p/MRP protein Pop1 and RNase p/MRP RNA components. RNA. 2015;21(9):1591–605.
4.
Zhu M, Wu C, Wu X, Song G, Li M, Wang Q. POP1 promotes the progression of breast cancer through maintaining telomere integrity. Carcinogenesis. 2023 Apr;44(3):252–62.
5.
Lemieux B, Laterreur N, Perederina A, Noël JF, Dubois ML, Krasilnikov AS, et al. Active yeast telomerase shares subunits with ribonucleoproteins RNase p and RNase MRP. Cell [Internet]. 2016;165(5):1171–81. Available from: https://www.sciencedirect.com/science/article/pii/S0092867416304093
6.
Rosenblad M, Lopez M, Piccinelli P, Samuelsson T. Inventory and analysis of the protein subunits of the ribonucleases p and MRP provides further evidence of homology between the yeast and human enzymes. Nucleic Acids Research [Internet]. 2006;34(18):5145–56. Available from: https://academic.oup.com/nar/article/34/18/5145/3112050
7.
Bateman A, Martin MJ, Orchard S, Magrane M, Ahmad S, Alpi E, et al. UniProt: The universal protein knowledgebase in 2023. Nucleic Acids Research. 2022 Nov;51(D1).
8.
Shakeel I, Zaidi Y, Gupta V, Badar S, Khan MKA, Abdullaev B, et al. Chapter 6 - emerging small-molecule inhibitors of ATM kinase targeting cancer therapy [Internet]. Hassan MdI, Noor S, editors. ScienceDirect. Academic Press; 2022. p. 179–98. Available from: https://www.sciencedirect.com/science/article/pii/B9780323912877000193
9.
Di Domenico EG, Romano E, Del Porto P, Ascenzioni F. Multifunctional role of ATM/Tel1 kinase in genome stability: From the DNA damage response to telomere maintenance. BioMed Research International. 2014;2014:1–17.
10.
Hu M, Zhou M, Bao X, Pan D, Jiao M, Liu X, et al. ATM inhibition enhances cancer immunotherapy by promoting mtDNA leakage and cGAS/STING activation. Journal of Clinical Investigation [Internet]. 2021 Feb;131(3). Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7843232/
11.
McKerlie M, Lin S, Zhu X-D. ATM regulates proteasome-dependent subnuclear localization of TRF1, which is important for telomere maintenance. Nucleic Acids Research. 2012 Jan;40(9):3975–89.
12.
Renault AL, Mebirouk N, Cavaciuti E, Le Gal D, Lecarpentier J, d’Enghien CD, et al. Telomere length, ATM mutation status and cancer risk in ataxia-telangiectasia families. Carcinogenesis [Internet]. 2017 Jul;38(10):994–1003. Available from: https://academic.oup.com/carcin/article/38/10/994/3994029
13.
Weber AM, Ryan AJ. ATM and ATR as therapeutic targets in cancer. Pharmacology & Therapeutics. 2015 May;149:124–38.
14.
Warren C, Pavletich NP. Structure of the human ATM kinase and mechanism of Nbs1 binding. eLife. 2022 Jan;11.
15.
Tong AS, Stern JL, Sfeir A, Kartawinata M, Lange T de, Zhu XD, et al. ATM and ATR signaling regulate the recruitment of human telomerase to telomeres. Cell Reports. 2015 Nov;13(8):1633–46.
16.
Hirano Y, Fukunaga K, Sugimoto K. Rif1 and Rif2 inhibit localization of Tel1 to DNA ends. Molecular Cell. 2009 Feb;33(3):312–22.
17.
Hector RE, Shtofman RL, Ray A, Chen BR, Nyun T, Berkner KL, et al. Tel1p preferentially associates with short telomeres to stimulate their elongation. Molecular Cell [Internet]. 2007 Sep;27(5):851–8. Available from: https://pubmed.ncbi.nlm.nih.gov/17803948/
18.
Fritz E, Friedl AA, Zwacka RM, Eckardt-Schupp F, Meyn MS. The yeast TEL1 gene partially substitutes for human ATM in suppressing hyperrecombination, radiation-induced apoptosis and telomere shortening in a-t cells. Molecular Biology of the Cell [Internet]. 2000 Aug;11(8):2605–16. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC14943/