Biology subjects

Mottet, D.

Publications and source records attributed to Mottet, D..

7 recordsLinked to original sources

bioRxiv2025

Unveiling the effect of phosphorylation on the structural and aggregation properties of the amyloidogenic intrinsically disordered protein DPF3a

The double plant homeodomain fingers 3a (DPF3 isoform a) is a human epigenetic regulator involved in chromatin remodelling, cell division, and ciliogenesis. Most notably, this protein is deregulated in various cancer types and neurodegenerative diseases. In our previous work, the disorder nature of DPF3a, as well as its propensity to aggregate into amyloid fibrils, have been highlighted, making it an amyloidogenic intrinsically disordered protein (IDP). Due to their high chain accessibility, IDPs structure and function are modulated by phosphorylation. It has been reported that phosphorylation of DPF3a at S348 (pS348) by the casein kinase 2 (CK2) is implicated in cardiac hypertrophy. CK2 can also phosphorylate DPF3a at S138 (pS138), which is also located in an intrinsically disordered region (IDR). However, no structural information is available on phosphorylated DPF3a. In the present study, we investigated the effect of phosphorylation on DPF3a structural and aggregation properties. Two single-mutated phosphomimetics (S138E and S348E) were characterised in vitro and compared to DPF3a WT, while in silico analyses were performed on pS138 and pS348 to assess structural changes at the molecular level. Circular dichroism and fluorescence spectroscopy revealed that both phosphomimetics are hybrid IDPs, with increased turn and antiparallel {beta}-sheet content as well as more buried aromatic residues compared to DPF3a WT, suggesting conformational rearrangements and a more folded N-terminal region. In silico characterisation supported these results, showing that phosphorylation of S138 and S348 induce extended conformation, especially the C-terminal extremity, due to electrostatic repulsion, while local folding occurs due to a proximity with arginine and lysine residues. Furthermore, spectroscopic and microscopic analyses unveiled that S138E and S348E exhibit slower fibrillation kinetics compared to DPF3a WT involving distinct aggregation mechanisms.

biophysics↗

bioRxiv2025

Exploration of the influence of environmental changes on the conformational and amyloidogenic landscapes of the zinc finger protein DPF3a by combining biophysical and molecular dynamics approaches

In the past few years, the double PHD fingers 3 (DPF3) protein isoforms (DPF3b and DPF3a) have been identified as new amyloidogenic intrinsically disordered proteins (IDPs). Although such discovery is coherent and promising in the light of their involvement in proteinopathies, their amyloidogenic pathway remains largely unexplored. As environmental variations in pH and ionic strength are relevant to DPF3 pathophysiological landscape, we therefore enquired the effect of these physicochemical parameters on the protein structural and prone-to-aggregation properties, by focusing on the more disordered DPF3a isoform. In the present study, we exploited in vitro and in silico strategies by combining spectroscopy, microscopy, and all-atom molecular dynamics methods. Very good consistency and complementary information were found between the experiments and the simulations. Acidification unequivocally abrogated DPF3a fibrillation upon maintaining the protein in highly hydrated and expanded conformers due to extensive repulsion between positively charged regions. In contrast, alkaline pH delayed the aggregation process due to loss in intramolecular contacts and chain decompaction, the extent of which was partly reduced thanks to the compensation of negative charge by arginine side chains. Through screening attractive electrostatic interactions, high ionic strength conditions (300 and 500 mM NaCl) shifted the conformational ensemble towards more swollen, heterogeneous, and less H-bonded structures, which were responsible for slowing down the conversion into {beta}-sheeted species and restricting the fibril elongation. For defining the self-assembly pathway of DPF3a, we unveiled that the protein amyloidogenicity intimately communicates with its conformational landscape, which is particularly sensitive to modification of its physicochemical environment. As such, understanding how to modulate DPF3a conformational ensemble will help designing novel protein-specific strategies for targeting neurodegeneration. HighlightsO_LIDPF3a is a polyampholyte IDP, structurally sensitive to environmental changes. C_LIO_LIDPF3a amyloid pathway and propensity can be modulated by pH and ionic strength. C_LIO_LIAcidic condition inhibits fibrillation and maintains DPF3a in an extended state. C_LIO_LIAlkaline pH and ionic strength delay fibrillation by reducing structure collapse. C_LIO_LIDPF3a fibrils exhibit condition-dependent optical-morphological properties. C_LI

biophysics↗

bioRxiv2024

Force Reserve Predicts Compensation in Reaching Movement with Induced Shoulder Strength Deficit

Following events such as fatigue or stroke, individuals often move their trunks forward during reaching, leveraging a broader muscle group even when only arm movement would suffice. In previous work, we showed the existence of a force reserve -- a phenomenon where individuals, when challenged with a heavy weight, adjusted their motor coordination to preserve approximately 40% of their shoulders force. Here, we investigated if such reserve can predict hip, shoulder, and elbow movements and torques resulting from an induced shoulder strength deficit. We engaged 20 healthy participants in a reaching task with incrementally heavier dumbbells, analyzing arm and trunk movements via motion capture and joint torques through inverse dynamics. We simulated these movements using an optimal control model of a 3-degree-of-freedom upper body, contrasting three cost functions: traditional sum of squared torques, a force reserve function incorporating a nonlinear penalty, and a normalized torque function. Our results demonstrate a clear increase in trunk movement correlated with heavier dumbbell weights, with participants employing compensatory movements to maintain a shoulder force reserve of approximately 40% of maximum torque. Simulations showed that while traditional and reserve functions accurately predicted trunk compensation, only the reserve function effectively predicted joint torques under heavier weights. These findings suggest that compensatory movements are strategically employed to minimize shoulder effort and distribute load across multiple joints in response to weakness. We discuss the implications of the force reserve cost function in the context of optimal control of human movements and its relevance for understanding of compensatory movements post-stroke. NEW & NOTEWORTHYOur study reveals key findings on compensatory movements during upper limb reaching tasks under shoulder strength deficits, as observed post-stroke. Using heavy dumbbells with healthy volunteers, we demonstrate how forward trunk displacement conserves around 40% of shoulder strength reserve during reaching. We show that an optimal controller employing a cost function combining squared motor torque and a nonlinear penalty for excessive muscle activation outperforms traditional controllers in predicting torques and compensatory movements in these scenarios.

neuroscience↗

bioRxiv2024

Intrinsic Disorder and Salt-dependent Conformational Changes of the N-terminal TFIP11 Splicing Factor

Tuftelin Interacting Protein 11 (TFIP11) was recently identified as a critical human spliceosome assembly regulator, interacting with multiple spliceosome proteins and localises in several membrane-less organelles. However, there is a lack of structural information on TFIP11, limiting the rationalisation of its biological role. TFIP11 has been predicted as a highly disordered protein, and more specifically concerning its N-terminal (N-TER) region. Intrinsically disordered proteins (IDPs) lack a defined tertiary structure, existing as a dynamic conformational ensemble, favouring their role as hubs in protein-protein and protein-RNA interaction networks. Furthermore, IDPs are involved in liquid-liquid phase separation (LLPS), driving the formation of subnuclear compartments. Combining disorder prediction, molecular dynamics, and spectroscopy methods, this contribution shows the first evidence TFIP11 N-TER may be described as a polyampholytic IDP, exhibiting a structural duality with the coexistence of ordered and disordered assemblies, depending on the ionic strength of the protein environment. Increasing the salt concentration enhances the protein conformational flexibility, presenting a fuzzier conformational landscape, a more globule-like shape, and an unstructured arrangement that could favour LLPS segregation and protein-RNA interaction. The regions mostly composed of charged and hydrophilic residues are the most impacted, including the G-Patch domain which is of crucial importance to TFIP11 function.

bioinformatics↗

bioRxiv2022

Validity and reliability of Kinect v2 for quantifying upper body kinematics during seated reaching

BackgroundKinematic analysis of the upper limbs is a good way to assess and monitor recovery in individuals with stroke, but remains little used in clinical routine due to its low feasibility. The aim of this study is to assess the validity and reliability of the Kinect v2 for the assessment of 17 kinematic variables commonly used in the analysis of upper limb reaching in stroke. Methods26 healthy participants performed seated hand-reaching tasks while holding a dumbbell to induce a behaviour similar to that of a person with a stroke. 3D upper limb and trunk motion were simultaneously recorded with the Kinect v2 (Microsoft, USA) and with the VICON (OxfordMetrics, UK), the latter being the reference system. For each kinematic outcome, the validity of the Kinect was assessed with ICC, linear regression and Bland & Altman plots. ResultsThe Kinect assesses trunk compensations, hand range of motion, movement time and mean velocity with a moderate to excellent reliability. In contrast, elbow and shoulder range of motion, time to peak velocity and path length ratio have a poor to moderate reliability, indicating that these variables should be interpreted with caution. Finally, instantaneous hand and elbow tracking are not precise enough to reliably assess Cartesian and angular kinematics over time, rendering variables such as the number of velocity peaks and the peak hand velocity unusable. ConclusionsThanks to its ease of use and markerless properties, the Kinect can be used in clinical routine for semi-automated quantitative diagnostics guiding individualised rehabilitation of the upper limb. However, engineers and therapists must bear in mind the limitations of the Kinect for the instantaneous tracking of the hand and elbow.

bioengineering↗

bioRxiv2021

The reserve of joint torque determines movement coordination

Humans coordinate biomechanical degrees of freedom to perform tasks at minimum cost. When reaching a target from a seated position, the trunk-arm-forearm coordination moves the hand to the well-defined spatial goal, while typically minimising hand jerk and trunk motion. However, due to fatigue or stroke, people visibly move the trunk more, and it is unclear what cost can account for this. Here we show that people recruit their trunk when the torque at the shoulder is too close to the maximum. We asked 26 healthy participants to reach a target while seated and we found that the trunk contribution to hand displacement increases from 11% to 27% when an additional load is handled. By flexing and rotating the trunk, participants spontaneously increase the reserve of anti-gravitational torque at the shoulder from 25% to 40% of maximal voluntary torque. Our findings provide hints on how to include the reserve of torque in the cost function of optimal control models of human coordination in healthy fatigued persons or in stroke victims.

neuroscience↗

bioRxiv2020

Proximal arm non-use optimises movement when the shoulder is weak: consequences for stroke patients

Most stroke patients do not use their paretic limb whereas they are able to. The Constraint-Induced Movement Therapy (CIMT) is effective to reverse this non-use behaviour in some patients but is inapplicable or unsuccessful on others. Here, we investigate how much non-use could come from shoulder weakness instead of the behavioural conditioning treated by the CIMT. We asked 26 healthy participants to reach a target while holding a dumbbell. We found that 18/26 participants exhibit proximal arm non-use when loaded and that non-use reduces shoulder torque of final posture. We either found that non-use improves accuracy in a high gravity field. Following optimal control policy, we explain how the non-use could be an adaptative solution when the shoulder is weak. Our results show the need to include muscular strength into cost function used to model human movement. The framework presented here suggests that psychological non-use could be treated effectively with CIMT, while physiological non-use, resulting from shoulder weakness, might respond better to anti-gravity muscles strengthening.

neuroscience↗