PMC4109683_01
Brain MRI examinations of Patient 1 (A-B) and Patient 2 (C-D), with corresponding normal control images (E-G) and muscle MRI examination of Patient 1 (H) and Patient 2 (I) at thigh and calf level. Axial A) and right parasagittal B) 3D SPGR T1-weighted images show abnormal gyration and delicate PMG-like cortical malformation involving the insulae (white arrowheads) and Heschle gyri (white arrows) with abnormal vertical orientation of the posterior part of sylvian fissure (black arrow). C) Axial 3D TFE T1-weighted image reveals bilateral perisylvian PMG-like cortical malformation involving the insulae (white arrowheads) and Heschle gyri (white arrows). Note an additional area of abnormal cortical gyration with infolding (black arrows) in the right occipital lobe. D) Coronal 3D TFE T1-weighted image depicts another area of abnormal cortical gyration with infolding (black arrow) in the right posterior cingulum slightly deforming the right lateral ventricle.. In muscle MRIs of lower limbs patients present diffuse atrophy and fat replacement of the thigh muscles which appear more prominent in Patient 2 (I), who accordingly displays the most severe phenotype. A selective similar sparing with compensatory hypertrophy of the adductor longus (al) and semitendinosus (st) muscles is evident to a similar extent in both cases. At leg level, MRI showed severe fat replacement of medial gastrocnemii (mg) with relative preservation of anterior tibialis (ta).
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F1
Proximal to the knee, the muscle scans displayed a pattern of diffuse atrophy and fat replacement of the thigh muscles, however there was selective hypertrophy of the adductor longus and semitendinosus muscles (Figure 1 H-I). Distal to the knee, the MRI showed preferential involvement of the posterior compartment with severe fat replacement of both gastrocnemii and relative preservation of anterior tibialis (Figure 1H-I). Figure S1 D) glutamic acid with lysine.
PMC4109683_02
Brain MRI examinations of Patient 1 (A-B) and Patient 2 (C-D), with corresponding normal control images (E-G) and muscle MRI examination of Patient 1 (H) and Patient 2 (I) at thigh and calf level. Axial A) and right parasagittal B) 3D SPGR T1-weighted images show abnormal gyration and delicate PMG-like cortical malformation involving the insulae (white arrowheads) and Heschle gyri (white arrows) with abnormal vertical orientation of the posterior part of sylvian fissure (black arrow). C) Axial 3D TFE T1-weighted image reveals bilateral perisylvian PMG-like cortical malformation involving the insulae (white arrowheads) and Heschle gyri (white arrows). Note an additional area of abnormal cortical gyration with infolding (black arrows) in the right occipital lobe. D) Coronal 3D TFE T1-weighted image depicts another area of abnormal cortical gyration with infolding (black arrow) in the right posterior cingulum slightly deforming the right lateral ventricle.. In muscle MRIs of lower limbs patients present diffuse atrophy and fat replacement of the thigh muscles which appear more prominent in Patient 2 (I), who accordingly displays the most severe phenotype. A selective similar sparing with compensatory hypertrophy of the adductor longus (al) and semitendinosus (st) muscles is evident to a similar extent in both cases. At leg level, MRI showed severe fat replacement of medial gastrocnemii (mg) with relative preservation of anterior tibialis (ta).
nihms598979f1.jpg
PMC4109683_03_nihms598979f1.jpg
F1
Proximal to the knee, the muscle scans displayed a pattern of diffuse atrophy and fat replacement of the thigh muscles, however there was selective hypertrophy of the adductor longus and semitendinosus muscles (Figure 1 H-I). Distal to the knee, the MRI showed preferential involvement of the posterior compartment with severe fat replacement of both gastrocnemii and relative preservation of anterior tibialis (Figure 1H-I). Figure S1 D) glutamic acid with lysine.
PMC4109683_03
Brain MRI examinations of Patient 1 (A-B) and Patient 2 (C-D), with corresponding normal control images (E-G) and muscle MRI examination of Patient 1 (H) and Patient 2 (I) at thigh and calf level. Axial A) and right parasagittal B) 3D SPGR T1-weighted images show abnormal gyration and delicate PMG-like cortical malformation involving the insulae (white arrowheads) and Heschle gyri (white arrows) with abnormal vertical orientation of the posterior part of sylvian fissure (black arrow). C) Axial 3D TFE T1-weighted image reveals bilateral perisylvian PMG-like cortical malformation involving the insulae (white arrowheads) and Heschle gyri (white arrows). Note an additional area of abnormal cortical gyration with infolding (black arrows) in the right occipital lobe. D) Coronal 3D TFE T1-weighted image depicts another area of abnormal cortical gyration with infolding (black arrow) in the right posterior cingulum slightly deforming the right lateral ventricle.. In muscle MRIs of lower limbs patients present diffuse atrophy and fat replacement of the thigh muscles which appear more prominent in Patient 2 (I), who accordingly displays the most severe phenotype. A selective similar sparing with compensatory hypertrophy of the adductor longus (al) and semitendinosus (st) muscles is evident to a similar extent in both cases. At leg level, MRI showed severe fat replacement of medial gastrocnemii (mg) with relative preservation of anterior tibialis (ta).
nihms598979f1.jpg
PMC4109683_03_nihms598979f1.jpg
F1
Proximal to the knee, the muscle scans displayed a pattern of diffuse atrophy and fat replacement of the thigh muscles, however there was selective hypertrophy of the adductor longus and semitendinosus muscles (Figure 1 H-I). Distal to the knee, the MRI showed preferential involvement of the posterior compartment with severe fat replacement of both gastrocnemii and relative preservation of anterior tibialis (Figure 1H-I). Figure S1 D) glutamic acid with lysine.
A-B Protein modeling. A) Cartoon representation of secondary structure elements of the tail/linker region of cytoplasmic dynein 1 heavy chain 1 protein. The model exhibits the typical three helix fold of the spectrin repeat-domain (pfam00435) with the. Gln1194 residue (highlighted in red) located in the alpha-helix B. The box is shows a magnification of the region of interest with the predicted consequences of the replacement of Arginine for Glutamine at residue 1194. The effect of mutation on protein structure and stability, as predicted by Rosetta Backrub server and FoldX, is that it determines a change in the conformation of the alpha-helix B and decreases the global stability of the domain (DeltaDeltaG: 4.66 +- 0.18 kcal/mol).. B) Cartoon (left) and electrostatic surface (right) representations of wild-type (upper) and c.9142G>A (p.Glu3048Lys) (lower) AAA4-alpha/beta submodule of human DYNC1H1 (residues 2870-3096). The AAA4-alpha/beta domain exhibits the Rossmann-type fold with the Glu3048 residue located in the pre-Sensor I region (colored in red), the characteristic insert that extends from alpha-helix 3 and beta-sheet 4 of the AAA4 module. The c.9142G>A (p.Glu3048Lys) mutation slightly decreases the global stability of the domain (DeltaDeltaG: 1.06+-0.08 kcal/mol) but severely alters the electrostatic surface of the pre-Sensor I region of AAA4 domain. Electrostatic surface potentials were colored according to charge with blue denoting positive charge (+5 kT/e-) and red, negative charge (-5 kT/e-).. C-E Nocodazole washout experiments with cultured skin fibroblasts from patients 1 and 2 (yellow and green bars respectively) or a control individual (cyan bars) lacking the dynein mutations. Results are shown as percentage of cells from each cell line with bracketed numbers representing the number of cells examined. C) For all cell lines, at the time of nocodazole exposure, nearly 100% of cells had a fully disrupted Golgi. D) At 30 minutes after nocodazole washout, a higher percentage of cells from patients 1 and 2, as compared to control cells, showed a fully disrupted Golgi. E) At 90 minutes after nocodazole washout, the percentage of cells with a fully disrupted Golgi was higher in patient cells compared to control cells. This was particularly evident in patient 2, where the number of cells retaining a fully disrupted Golgi was nearly 50% of the total. Results are taken from at least 3 Independent experiments. Error bars=standard deviation; n= number of cells. Unpaired t-test: *, p<0.05 **, p<0.001.. F. Representative images of cultured skin fibroblasts with intact (upper panel), partially disrupted (middle panel) and fully disrupted (lower panel) Golgi are shown. Golgi is stained in red. Scale bars=5mum.
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Methods), suggested that p.Q1194R alters the conformation of the region lowering the global stability of this domain (Figure 2A). AAA4 module, but might decrease the global stability of the domain since the mutation drastically changes electrostatic surface potential of the entire module (Figure 2B). According to the crystal structures of dynein, the pre-Sensor The effect was more severe in Patient 2 than in Patient 1 (Figure 2C-E). Figure S2). Figure S2).