Self-oligomerization regulates stability of survival motor neuron protein isoforms by sequestering an SCF^Slmb degron

Spinal muscular atrophy (SMA) is caused by homozygous mutations in human SMN1. Expression of a duplicate gene (SMN2) primarily results in skipping of exon 7 and production of an unstable protein isoform, SMNΔ7. Although SMN2 exon skipping is the principal contributor to SMA severity, mechanisms governing stability of survival motor neuron (SMN) isoforms are poorly understood. We used a Drosophila model system and label-free proteomics to identify the SCFSlmb ubiquitin E3 ligase complex as a novel SMN binding partner. SCFSlmb interacts with a phosphor degron embedded within the human and fruitfly SMN YG-box oligomerization domains. Substitution of a conserved serine (S270A) interferes with SCFSlmb binding and stabilizes SMNΔ7. SMA-causing missense mutations that block multimerization of full-length SMN are also stabilized in the degron mutant background. Overexpression of SMNΔ7S270A, but not wild-type (WT) SMNΔ7, provides a protective effect in SMA model mice and human motor neuron cell culture systems. Our findings support a model wherein the degron is exposed when SMN is monomeric and sequestered when SMN forms higher-order multimers

Synergistic interplay of Gβγ and phosphatidylinositol 4,5-bisphosphate dictates Kv7.4 channel activity.

Kv7.4 channels are key determinants of arterial contractility and cochlear mechanosensation that, like all Kv7 channels, have an obligatory requirement for phosphatidylinositol 4,5-bisphosphate (PIP2). βγ G proteins (Gβγ) have been identified as novel positive regulators of Kv7.4. The present study ascertained whether Gβγ increased Kv7.4 open probability through an increased sensitivity to PIP2. In HEK cells stably expressing Kv7.4, PIP2 or Gβγ increased open probability in a concentration dependent manner. Depleting PIP2 prevented any Gβγ-mediated stimulation whilst an array of Gβγ inhibitors prohibited any PIP2-induced current enhancement. A combination of PIP2 and Gβγ at sub-efficacious concentrations increased channel open probability considerably. The stimulatory effects of three Kv7.2-7.5 channel activators were also lost by PIP2 depletion or Gβγ inhibitors. This study alters substantially our understanding of the fundamental processes that dictate Kv7.4 activity, revealing a more complex and subtle paradigm where the reliance on local phosphoinositide is dictated by interaction with Gβγ

Effects of barefoot and shod running on lower extremity joint loading, a musculoskeletal simulation study

PURPOSE: The aim of the current investigation was to utilize a musculoskeletal simulation based approach, to examine the effects of barefoot and shod running on lower extremity joint loading during the stance phase. METHODS: Twelve male runners, ran over an embedded force plate at 4.0 m/s, in both barefoot and shod conditions. Kinematics of the lower extremities were collected using an eight camera motion capture system. Lower extremity joint loading was also explored using a musculoskeletal simulation and mathematical modelling approach, and differences between footwear conditions were examined using paired samples t-tests. RESULTS: Peak Achilles tendon force was significantly larger (P=0.039) when running barefoot (6.85 BW) compared to shod (6.07 BW). In addition, both medial (P=0.013) and lateral (P=0.007) tibiofemoral instantaneous load rates were significantly larger in the barefoot (medial = 289.17 BW/s & lateral = 179.59 BW/s) in relation to the shod (medial = 167.57 BW/s & lateral = 116.40 BW/s) condition. Finally, the barefoot condition (9.70 BW) was associated with a significantly larger (P=0.037) peak hip force compared to running shod (8.51 BW). CONCLUSIONS: The current investigation indicates that running barefoot may place runners at increased risk from the biomechanical factors linked to the aetiology of chronic lower extremity pathologies. However, future analyses using habitual barefoot runners, are required before more definitive affirmations regarding injury predisposition can be made