Structure-Guided Exploration of SDS22 Interactions with Protein Phosphatase PP1 and the Splicing Factor BCLAF1

Heroes, Ewald; Hoeven, Gerd Van der; Choy, M.S.; García, Javier del Pino; Ferreira, Mónica; Nys, Mieke; Derua, Rita; Beullens, Monique; Ulens, Chris; Peti, Wolfgang; Meervelt, Luc Van; Page, Rebecca

doi:10.1016/j.str.2018.12.002

Cited by 20 publications

(37 citation statements)

References 68 publications

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“…These data are consistent with our PP1 Y134A SPR data ( Fig. 4D) and previously published PP1 K147A/K150A pull-down data (37) which also show that these variants fail to bind SDS22. Together, these data show that, both in vivo and in vitro, Lys147/ Lys150 and Tyr134 are critical for SDS22 binding.…”

Section: Sds22 and Other Pp1 Regulatory Proteins Prevent Pp1 Aggregatsupporting

confidence: 94%

“…2A). No electron density was observed for SDS22 residues 56-75, consistent with previous data demonstrating that these residues are both dispensable for PP1 binding (22) and less well ordered than the Leu-rich repeats (LRRs) (36,37). The SDS22:PP1 interaction surface is extensive, burying more than 3084 Å 2 of solvent accessible surface area (Fig.…”

Section: Sds22 and Other Pp1 Regulatory Proteins Prevent Pp1 Aggregatsupporting

confidence: 87%

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SDS22 selectively recognizes and traps metal-deficient inactive PP1

Choy

Moon

Ravindran

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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The metalloenzyme protein phosphatase 1 (PP1), which is responsible for ≥50% of all dephosphorylation reactions, is regulated by scores of regulatory proteins, including the highly conserved SDS22 protein. SDS22 has numerous diverse functions, surprisingly acting as both a PP1 inhibitor and as an activator. Here, we integrate cellular, biophysical, and crystallographic studies to address this conundrum. We discovered that SDS22 selectively binds a unique conformation of PP1 that contains a single metal (M2) at its active site, i.e., SDS22 traps metal-deficient inactive PP1. Furthermore, we showed that SDS22 dissociation is accompanied by a second metal (M1) being loaded into PP1, as free metal cannot dissociate the complex and M1-deficient mutants remain constitutively trapped by SDS22. Together, our findings reveal that M1 metal loading and loss are essential for PP1 regulation in cells, which has broad implications for PP1 maturation, activity, and holoenzyme subunit exchange.

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Section: Sds22 and Other Pp1 Regulatory Proteins Prevent Pp1 Aggregatsupporting

confidence: 94%

Section: Sds22 and Other Pp1 Regulatory Proteins Prevent Pp1 Aggregatsupporting

confidence: 87%

SDS22 selectively recognizes and traps metal-deficient inactive PP1

Choy

Moon

Ravindran

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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“…Consistent with these reports, our results indicated that BCLAF1 knockdown reduced cell proliferation and increased cell susceptible to IR-induced DNA damages in GC cells. Recent studies indicated that BCLAF1 function mainly depends on its phosphorylation, for example, Ewald Heroes et al reported that the BCLAF1:SDS22 interaction is dependent on the phosphorylation of BCLAF1 [29]. By searching the uniport protein database, 46 phosphorylation sites were found in BCLAF1, but the signi cance and function of these sites have not been reported.…”

Section: Discussionmentioning

confidence: 99%

Bcl-2-associated Transcription Factor 1 Phosphorylation Mediated DNA Damage Response in Gastric Cancer

Liu

Sun

et al. 2021

Preprint

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Background: DNA damage response plays critical roles in tumor pathogenesis and radiotherapy resistance. Various cellular activities including the initiation and major steps DNA damage response are regulated by protein phosphorylation, whereas the molecular mechanism remains largely unknown. Methods: A quick label-free phosphoproteomics using high-resolution mass spectrometry and an open search approach was applied to paired tumor and adjacent tissues from five patients with gastric cancer. The dysregulated phosphoproteins were identified and their associated-pathways analyzed using Gene Set Enrichment Analysis (GSEA). The mostly regulated phosphoproteins and their potential functions were validated by the specific antibodies against the phosphorylation sites. Specific protein phosphorylation was further analyzed by functional and clinical approaches. Results: 832 gastric cancer-associated unique phosphorylated sites were identified, among which 25 were up- and 52 down-regulated. Markedly, the dysregulated phosphoproteins were primarily enriched in DNA-damage-response-associated pathways. Particularly, the phosphorylation of Bcl-2-associated transcription factor 1 (BCLAF1) at Ser290 was significantly upregulated in tumor. The upregulation of BCLAF1 Ser290 phosphorylation (pBCLAF1(Ser290)) in tumor was confirmed by tissue microarray studies and further indicated in association with poor prognosis of gastric cancer patients. Eliminating the phosphorylation of BCLAF1 at Ser290 suppressed gastric cancer (GC) cell proliferation. Upregulation of pBCLAF1(Ser290) was found in association with irradiation-induced γ-H2AX expression in the nucleus, leading to an increased DNA damage repair response, and a marked inhibition of irradiation-induced cancer cell apoptosis. Conclusions: The phosphorylation of BCLAF1 at Ser290 is involved in the regulation of DNA damage response, indicating an important target for the resistance of radiotherapy.

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“…Apart from LRR-containing genes from human pathogen Leptospira interrogans such as LIC11098 that exhibits a high structural similarity with Zscores of 26.6 (root-mean-square-deviation, RMSD, of 2.0 Å) (24), it is noteworthy that the structure of Rsu-1 resembles that of a human protein phosphatase 1 regulatory subunit 7 (a.k.a. SDS22) (25), a prototype of SDS22-like subfamily in the LRR superfamily (10) (Figure S1). Rsu-1 shares a 27% sequence identity with SDS22 that comprises 12 LRRs (25).…”

mentioning

confidence: 99%

“…SDS22) (25), a prototype of SDS22-like subfamily in the LRR superfamily (10) (Figure S1). Rsu-1 shares a 27% sequence identity with SDS22 that comprises 12 LRRs (25). The structure of Rsu-1 superimposes with that of SDS22 with an RMSD of 2.8 Å for the equivalent C atom pairs.…”

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confidence: 99%

Molecular basis for Ras suppressor-1 recruitment to focal adhesions and stabilization of consensus adhesome complex

Fukuda

Lü

Qin

2021

Preprint

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Ras suppressor-1 (Rsu-1) is a leucine-rich repeat (LRR)-containing protein that is crucial for regulating fundamental cell adhesion processes and tumor development. Rsu-1 interacts with a zinc-finger type multi LIM domain-containing adaptor protein PINCH-1 involved in the integrin-mediated consensus adhesome but not with highly homologous isoform PINCH-2. However, the structural basis for such specific interaction and regulatory mechanism remains unclear. Here, we determined the crystal structures of Rsu-1 and its complex with the PINCH-1 LIM4-5 domains. Rsu-1 displays an arc-shaped solenoid architecture with eight LRRs shielded by the N- and C-terminal capping modules. We show that a large conserved concave surface of the Rsu-1 LRR domain recognizes the PINCH-1 LIM5 domain, and that the C-terminal non-LIM region of PINCH-2 but not PINCH-1 sterically disfavors the Rsu-1 binding. We further show that Rsu-1 can be assembled, via PINCH-1-binding, into a tight hetero-pentamer complex comprising Rsu-1, PINCH-1, ILK, Parvin, and Kindlin-2 that constitute a major consensus integrin adhesome crucial for focal adhesion assembly. Consistently, our mutagenesis and cell biological data consolidate the significance of the Rsu-1/PINCH-1 interaction in focal adhesion assembly and cell spreading. Our results provide a crucial molecular insight into Rsu-1-mediated cell adhesion with implication on how it may regulate tumorigenic growth.

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Structure-Guided Exploration of SDS22 Interactions with Protein Phosphatase PP1 and the Splicing Factor BCLAF1

Abstract: Highlights d The LRRs of SDS22 interact with PP1 helices a5 and a6 d A basic surface patch of SDS22 binds phosphorylated BCLAF1 d Many PP1 holoenzymes contain SDS22 as a third subunit d SDS22 prevents the autodephosphorylation of PP1

Cited by 20 publications

References 68 publications

SDS22 selectively recognizes and traps metal-deficient inactive PP1

SDS22 selectively recognizes and traps metal-deficient inactive PP1

Bcl-2-associated Transcription Factor 1 Phosphorylation Mediated DNA Damage Response in Gastric Cancer

Molecular basis for Ras suppressor-1 recruitment to focal adhesions and stabilization of consensus adhesome complex

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