Structural basis of the human Scribble-Vangl2 association in health and disease

How, Jing Yuan; Stephens, Rebecca; Lim, Krystle Y.B.; Humbert, Patrick O.; Kvansakul, Marc

doi:10.1101/2020.10.07.330712

Cited by 3 publications

(4 citation statements)

References 46 publications

(72 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Lastly, the conserved His in each domain found on a2, PDZ1 H793, PDZ2 H928, and PDZ3 H1071, shared a hydrogen bond with Thr351 Tax1 , and mutation to Ala ablates Tax1 binding to SCRIB PDZ domains as seen in our ITC analysis (Table 1, Figs 1B and 3). This particular conserved His residue is also involved in interactions with other cellular and viral interactors of Scribble as observed in crystal structures of SCRIB PDZ1:b-PIX, PDZ3:b-PIX, PDZ1:MCC, PDZ1:APC, PDZ1:Vangl2, PDZ2: Vangl2, PDZ3:Vangl2, PDZ1: NS1, and PDZ3:NS1 [20,37,[43][44][45], thus making this residue vital for SCRIB:ligand interactions.…”

Section: Discussionmentioning

confidence: 99%

“…The SCRIB PDZ2:Tax1 complex formed single ovoid disc shaped crystals belong to space group P6 1 22 with a = 100.77 Å, b = 100.77 Å, c = 68.53 Å, α = 90.00°, β = 90.00°, and γ = 120.00° in the hexagonal crystal system. Molecular replacement was carried out using PHASER [67] with the previously solved structure of SCRIB PDZ2 [45] as a search model. SCRIB PDZ2:Tax1 crystals contained 2 molecules of PDZ2 and 1 molecule of Tax1 peptide in the asymmetric unit, with 42.82% solvent content and final TFZ and LLG values of 23.1 and 507.5, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Human Scribble (SCRIB) PDZ1 (728-815), PDZ2 (860-950), PDZ3 (1002-1094), and PDZ4 (1099-1203) domain proteins (Uniprot accession number: Q14160) as well as mutant SCRIB PDZ domain proteins were purified as previously described [37,45]. SCRIB PDZ mutants cDNA PDZ3 H1071A (residues 1002-1094), PDZ4 R1110D (residues 1099-1203), PDZ4 R1116D (residues 1099-1203), and PDZ4 H1170A (residues 1099-1203) was cloned into the pGex-6P3 vector (Bioneer).…”

Section: Protein Expression and Purificationmentioning

confidence: 99%

See 2 more Smart Citations

Structural insight into the Scribble PDZ domains interaction with the oncogenic Human T‐cell lymphotrophic virus‐1 (HTLV‐1) Tax1 PBM

et al. 2022

Self Cite

View full text Add to dashboard Cite

Scribble (Scrib) is a highly conserved cell polarity regulator that harbours potent tumour suppressor activity and plays an important role in cell migration. Dysregulation of polarity is associated with poor prognosis during viral infections. Human T‐cell lymphotrophic virus‐1 (HTLV‐1) encodes for the oncogenic Tax1 protein, a modulator of the transcription of viral and human proteins that can cause cell cycle dysregulation as well as a loss of genomic integrity. Previous studies established that Scribble interacts with Tax1 via its C‐terminal PDZ‐binding motif (PBM), leading to aggregation of polarity regulators and subsequent perturbation of host cell adhesion, proliferation, and signalling. Using isothermal titration calorimetry, we now show that all four PDZ domains of Scribble bind to Tax1 PBM. We then determined crystal structures of Scribble PDZ1, PDZ2 and PDZ3 domains bound to Tax1 PBM. Our findings establish a structural basis for Tax1‐mediated subversion of Scribble‐mediated cell polarity signalling and provide the platform for mechanistic studies to examine Tax1 induced mislocalization of Scribble and the associated changes in cellular architecture and subsequent tumorigenesis.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Protein Expression and Purificationmentioning

confidence: 99%

See 1 more Smart Citation

Structural insight into the Scribble PDZ domains interaction with the oncogenic Human T‐cell lymphotrophic virus‐1 (HTLV‐1) Tax1 PBM

et al. 2022

Self Cite

View full text Add to dashboard Cite

show abstract

“…Raw thermograms were processed with MicroCal Origin® version 7.0 software (OriginLabTM Corporation) to obtain the binding parameters of each interaction. A synthetic pan-PDZ binding peptide referred to as superpeptide (RSWFETWV) was used as a positive control [42].…”

Section: Isothermal Titration Calorimetrymentioning

confidence: 99%

Structural analysis of human papillomavirus E6 interactions with Scribble PDZ domains

Stewart

Caria

Humbert

et al. 2022

Preprint

View full text Add to dashboard Cite

The cell polarity regulator Scribble has been shown to be a critical regulator of the establishment and development of tissue architecture, and its dysregulation promotes or suppresses tumor development in a context dependent manner. Scribble activity is subverted by numerous viruses. This includes human papillomaviruses (HPVs), who target Scribble via the E6 protein. Binding of E6 from high-risk HPV strains to Scribble via a C-terminal PDZ binding motif leads to Scribble degradation in vivo. However, the precise molecular basis for Scribble-E6 interactions remains to be defined. We now show that Scribble PDZ1 and PDZ3 are the major interactors of HPV E6 from multiple high-risk strains, with each E6 protein displaying a unique interaction profile. We then determined crystal structures of Scribble PDZ1 and PDZ3 domains in complex with the PBM motifs of E6 from HPV strains 16, 18 and 66. Our findings reveal distinct interaction patterns for each E6 PBM motif from a given HPV strain, suggesting that a complex molecular interplay exists that underpins the overt Scribble-HPV E6 interaction and controls E6 carcinogenic potential.

show abstract

APOLR3B-variant reveals a Pol III transcriptome response dependent on La protein/SSB

Mattijssen,

Kerkhofs,

Stephen

et al. 2024

Preprint

View full text Add to dashboard Cite

RNA polymerase III (Pol III, POLR3) synthesizes tRNAs and other small non-coding RNAs. HumanPOLR3pathogenic variants cause a range of developmental disorders, recapitulated in part by mouse models, yet some aspects of POLR3 deficiency have not been explored. We characterized a humanPOLR3B:c.1625A>G;p.(Asn542Ser) disease variant that was found to cause mis-splicing ofPOLR3B. Genome-editedPOLR3B1625A>GHEK293 cells acquired the mis-splicing with decreases in multiple POLR3 subunits and TFIIIB, although display auto-upregulation of the Pol III termination-reinitiation subunitPOLR3E. La protein was increased relative to its abundant pre-tRNA ligands which bind via their U(n)U-3’-termini. Assays for cellular transcription revealed greater deficiencies for tRNA genes bearing terminators comprised of 4Ts than of ≥5Ts. La-knockdown decreased Pol III ncRNA expression unlinked to RNA stability. Consistent with these effects, small-RNAseq showed thatPOLR3B1625A>Gand patient fibroblasts express more tRNA fragments (tRFs) derived from pre-tRNA 3’-trailers (tRF-1) than from mature-tRFs, and higher levels of multiple miRNAs, relative to control cells. The data indicate that decreased levels of Pol III transcripts can lead to functional excess of La protein which reshapes small ncRNA profiles revealing new depth in the Pol III system. Finally, patient cell RNA analysis uncovered a strategy for tRF-1/tRF-3 asPOLR3-deficiency biomarkers.

show abstract

Structural basis of the human Scribble-Vangl2 association in health and disease

Cited by 3 publications

References 46 publications

Structural insight into the Scribble PDZ domains interaction with the oncogenic Human T‐cell lymphotrophic virus‐1 (HTLV‐1) Tax1 PBM

Structural insight into the Scribble PDZ domains interaction with the oncogenic Human T‐cell lymphotrophic virus‐1 (HTLV‐1) Tax1 PBM

Structural analysis of human papillomavirus E6 interactions with Scribble PDZ domains

APOLR3B-variant reveals a Pol III transcriptome response dependent on La protein/SSB

Contact Info

Product

Resources

About