Molecular basis for substrate recruitment to the PRMT5 methylosome

Mulvaney, Kathleen M.; Blomquist, Christa; Acharya, Nischal; Li, Ruitong; O’Keefe, Meghan; Ranaghan, Matthew J.; Stokes, Matthew P.; Nelson, Alissa J.; Jain, Sidharth; Columbus, Josie; Bozal, Fazli K.; Skepner, Adam; Raymond, D.D.; McKinney, David C.; Freyzon, Yelena; Baidi, Yossef; Ianari, Alessandra; McMillan, Brian J.; Sellers, William R.

doi:10.1101/2020.08.22.256347

Cited by 5 publications

(18 citation statements)

References 49 publications

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“…A co-crystal structure of a RioK1 derived peptide with the PRMT5 TIM barrel domain confirmed the results of our mutation and truncation studies. Our findings are in agreement with the recent preliminary report by Sellers et al [7]…”

Section: Introductionsupporting

confidence: 94%

“…). [7] symmetry molecule, while the N-terminal part (Ser9, Arg10 and Val11) is strongly solvent exposed. The core of the sequence (GQFDDAD), however, is forming significant interactions with the protein groove.…”

Section: Resultsmentioning

confidence: 99%

“…[3a,c] PRMT5 and MEP50 form a heterooctameric complex whose structure has been determined, [3a] and structural information on the interaction between PRMT5 and the other adaptor proteins has only very recently been described in preliminary form. [7] Overexpression of PRMT5 is frequently observed in cancer which makes the protein an intensively investigated anticancer target, and active-site directed inhibitors have reached clinical trials. [8] Further study of the PRMT5 PPIs and binding partners is important for a better understanding of its regulation.…”

Section: Introductionmentioning

confidence: 99%

“…D) Close-up of 23 bound to the TIM barrel. E) Superposition of the RioK1derived peptides from PDB structures 7BOC (grey, this work) and 6V0N (yellow, Sellers et al) [7].…”

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confidence: 99%

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Biochemical Investigation of the Interaction of pICln, RioK1 and COPR5 with the PRMT5–MEP50 Complex

et al. 2021

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The PRMT5–MEP50 methyltransferase complex plays a key role in various cancers and is regulated by different protein–protein interactions. Several proteins have been reported to act as adaptor proteins that recruit substrate proteins to the active site of PRMT5 for the methylation of arginine residues. To define the interaction between these adaptor proteins and PRMT5, we employed peptide truncation and mutation studies and prepared truncated protein constructs. We report the characterisation of the interface between the TIM barrel of PRMT5 and the adaptor proteins pICln, RioK1 and COPR5, and identify the consensus amino acid sequence GQF[D/E]DA[E/D] involved in binding. Protein crystallography revealed that the RioK1 derived peptide interacts with a novel PPI site.

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Section: Introductionsupporting

confidence: 94%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…D) Close-up of 23 bound to the TIM barrel. E) Superposition of the RioK1derived peptides from PDB structures 7BOC (grey, this work) and 6V0N (yellow, Sellers et al) [7].…”

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confidence: 99%

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Biochemical Investigation of the Interaction of pICln, RioK1 and COPR5 with the PRMT5–MEP50 Complex

et al. 2021

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“…(D) Left panel: WB analysis of total symmetric dimethylation levels in MTAP-/-HCT116 cells, in response to BRD0639 or BRD2198. Right panel: WB analysis of total symmetric dimethylation levels in MTAP-/-HCT116 cells overexpressing the PRMT5 ADA mutant which is unable to bind PBM peptide(Mulvaney, 2020), with and without KO of endogenous PRMT5.Contributions A.I., D.C.M. and B.J.M.…”

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confidence: 99%

Discovery of a first-in-class inhibitor of the PRMT5-substrate adaptor interaction

McKinney

McMillan

Ranaghan

et al. 2021

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PRMT5 and its substrate adaptor proteins (SAPs), pICln and Riok1, are synthetic lethal dependencies in MTAP-deleted cancer cells. SAPs share a conserved PRMT5 binding motif (PBM) which mediates binding to a surface of PRMT5 distal to the catalytic site. This interaction is required for methylation of several PRMT5 substrates, including histone and spliceosome complexes. We screened for small molecule inhibitors of the PRMT5-PBM interaction and validated a compound series which binds to the PRMT5-PBM interface and directly inhibits binding of SAPs. Mode of action and structure determination studies revealed that these compounds form a covalent bond between a halogenated pyridazinone group and cysteine 278 of PRMT5. Optimization of the starting hit produced a lead compound, BRD0639, which engages the target in cells, disrupts the PRMT5-RIOK1 complex, and reduces substrate methylation. BRD0639 is a first-in-class PBM-competitive small molecule that can support studies of PBM-dependent PRMT5 activities and the development of novel PRMT5 inhibitors that selectively target these functions.

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Discovery of a First-in-Class Inhibitor of the PRMT5–Substrate Adaptor Interaction

et al. 2021

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PRMT5 and its substrate adaptor proteins (SAPs), pICln and Riok1, are synthetic lethal dependencies in MTAP-deleted cancer cells. SAPs share a conserved PRMT5 binding motif (PBM) which mediates binding to a surface of PRMT5 distal to the catalytic site. This interaction is required for methylation of several PRMT5 substrates, including histone and spliceosome complexes. We screened for small molecule inhibitors of the PRMT5–PBM interaction and validated a compound series which binds to the PRMT5–PBM interface and directly inhibits binding of SAPs. Mode of action studies revealed the formation of a covalent bond between a halogenated pyridazinone group and cysteine 278 of PRMT5. Optimization of the starting hit produced a lead compound, BRD0639, which engages the target in cells, disrupts PRMT5–RIOK1 complexes, and reduces substrate methylation. BRD0639 is a first-in-class PBM-competitive inhibitor that can support studies of PBM-dependent PRMT5 activities and the development of novel PRMT5 inhibitors that selectively target these functions.

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Molecular basis for substrate recruitment to the PRMT5 methylosome

Cited by 5 publications

References 49 publications

Biochemical Investigation of the Interaction of pICln, RioK1 and COPR5 with the PRMT5–MEP50 Complex

Biochemical Investigation of the Interaction of pICln, RioK1 and COPR5 with the PRMT5–MEP50 Complex

Discovery of a first-in-class inhibitor of the PRMT5-substrate adaptor interaction

Discovery of a First-in-Class Inhibitor of the PRMT5–Substrate Adaptor Interaction

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