Clinical, Genetics, and Bioinformatic Characterization of Mutations Affecting an Essential Region of PLS3 in Patients with BMND18

Figures

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  Figure 1. Genetic characterization of the patient described. (a) Family pedigree. Mutation status of PLS3 c.745G > T is indicated beneath symbols for each subject. WT indicates wild-type. (b) Schematic representation of the mapped exome sequencing reads visualized using the Integrative Genomics Viewer (IGV) browser for the proband and his parents. The red lines indicate the mutation. (c) Sanger sequencing of three individuals in the family over the c.745 position in PLS3. The upper panel shows the c.745G > T variant in the patient, the middle panel shows c.745G > T mutation in the heterozygous mother, and the lower panel shows WT in the patient's father.

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  Figure 2. Sequence conservation in the plastin-3 family. (a) Local view of the multiple sequence alignment of plastin-3 in the neighbourhood of Glu249 (red arrow). (b) Whole plastin-3 plot. Conservation is represented using Shannon entropy, a parameter routinely used for scoring pathogenic mutations, whose values range from 0 (complete conservation) to 4.32 (all amino acids are equiprobable at that position).

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  Figure 3. Structures of wild-type and mutant plastin-3. The run of the protein chain is shown as a secondary structure cartoon color-coded according to five domains. (a) Classical view of the structure of the wild-type plastin-3. The EF-hand domains are shown in yellow, CH1 in blue, CH2 in cyan, CH3 in green, CH4 in pink, and the LOOP-1 region in red, respectively. Mutant plastin-3 structures are shown in (b) (c.759_760 ins AAT), (c) (c.748ins36), (d) (c.1103C > A), and (e) (c.1433G > A), respectively. The location of the mutated residues is highlighted in black circle, and residues are highlighted in sticks.

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  Figure 4. The RMSD is calculated on Cα atoms of the wild-type plastin-3 (red) and the mutant (Glu249_Ala250ins12) (blue) structure (a), respectively. (b) Atomic fluctuations (RMSF) is calculated from the Cα of the wild-type (red) and mutant (blue) plastin-3 during MD simulations.

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  Figure 5. The hydrogen bonds formed between the CH2 and CH3 interface for wild plastin-3 system (a) and Glu249_Ala250ins12 system (b), respectively. The protein is represented with colored cartoon, red for the LOOP-1 region, blue for CH2 domain, and green for CH3 domain, respectively. Residues involved in the h-bonds interactions are highlighted with sticks.