CDKL5 and Shootin1 Interact and Concur in Regulating Neuronal Polarization

PLoS One. 2016 Feb 5;11(2):e0148634. doi: 10.1371/journal.pone.0148634. eCollection 2016.


In the last years, the X-linked cyclin-dependent kinase-like 5 (CDKL5) gene has been associated with epileptic encephalopathies characterized by the early onset of intractable epilepsy, severe developmental delay, autistic features, and often the development of Rett syndrome-like features. Still, the role of CDKL5 in neuronal functions is not fully understood. By way of a yeast two hybrid screening we identified the interaction of CDKL5 with shootin1, a brain specific protein acting as a determinant of axon formation during neuronal polarization. We found evidence that CDKL5 is involved, at least in part, in regulating neuronal polarization through its interaction with shootin1. Indeed, the two proteins interact in vivo and both are localized in the distal tip of outgrowing axons. By using primary hippocampal neurons as model system we find that adequate CDKL5 levels are required for axon specification. In fact, a significant number of neurons overexpressing CDKL5 is characterized by supernumerary axons, while the silencing of CDKL5 disrupts neuronal polarization. Interestingly, shootin1 phosphorylation is reduced in neurons silenced for CDKL5 suggesting that the kinase affects, directly or indirectly, the post-translational modification of shootin1. Finally, we find that the capacity of CDKL5 to generate surplus axons is attenuated in neurons with reduced shootin1 levels, in agreement with the notion that two proteins act in a common pathway. Altogether, these results point to a role of CDKL5 in the early steps of neuronal differentiation that can be explained, at least in part, by its association with shootin1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axons / metabolism*
  • Cell Polarity / physiology*
  • Hippocampus / cytology
  • Hippocampus / metabolism*
  • Mice
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism*
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism*


  • Nerve Tissue Proteins
  • Protein Serine-Threonine Kinases
  • CDKL5 protein, mouse

Grant support

This work was supported by Telethon (GGP10032), Ministero della Salute (Ricerca finalizzata 2008 – Bando Malattie Rare) and Cariplo (2010-0724) Foundations (to NL) and by the EU Initial Training Network program (no. 238242 “DisChrom”) and the Jerome Lejeune Foundation (to CKN), and the Italian parents’ associations L’Albero di Greta and ProRett Ricerca.