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A C-terminal motif containing a PKC phosphorylation site regulates γ-Protocadherin-mediated dendrite arborization in the cerebral cortex in vivo
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A C-terminal motif containing a PKC phosphorylation site regulates γ-Protocadherin-mediated dendrite arborization in the cerebral cortex in vivo

Camille M Hanes, Kar Men Mah, David M Steffen, Charles G Marcucci, Leah C Fuller, Robert W Burgess, Andrew M Garrett and Joshua A Weiner
bioRxiv : the preprint server for biology
Cold Spring Harbor Laboratory
01/25/2024
DOI: 10.1101/2024.01.25.577214
PMCID: PMC10849722
PMID: 38328061
url
https://doi.org/10.1101/2024.01.25.577214View
Preprint (Author's original)This preprint has not been evaluated by subject experts through peer review. Preprints may undergo extensive changes and/or become peer-reviewed journal articles. Open Access

Abstract

The gene cluster encodes 22 γ-Protocadherin (γ-Pcdh) cell adhesion molecules that critically regulate multiple aspects of neural development, including neuronal survival, dendritic and axonal arborization, and synapse formation and maturation. Each γ-Pcdh isoform has unique protein domains-a homophilically-interacting extracellular domain and a juxtamembrane cytoplasmic domain- as well as a C-terminal cytoplasmic domain shared by all isoforms. The extent to which isoform-specific shared domains regulate distinct γ-Pcdh functions remains incompletely understood. Our previous studies identified PKC phosphorylation of a serine residue within a shared C-terminal motif as a mechanism through which γ-Pcdh promotion of dendrite arborization MARCKS is abrogated. Here, we used CRISPR/Cas9 genome editing to generate two new mouse lines expressing only non-phosphorylatable γ-Pcdhs, due either to a serine-to-alanine mutation ( ) or to a 15-amino acid C-terminal deletion resulting from insertion of an early stop codon ( ). Both lines are viable and fertile, and the density and maturation of dendritic spines remains unchanged in both and cortex. Dendrite arborization of cortical pyramidal neurons, however, is significantly increased in both lines, as are levels of active MARCKS. Intriguingly, despite having significantly reduced levels of γ-Pcdh proteins, the mutation yields the strongest phenotype, with even heterozygous mutants exhibiting increased arborization. The present study confirms that phosphorylation of a shared C-terminal motif is a key γ-Pcdh negative regulation point, and contributes to a converging understanding of γ-Pcdh family function in which distinct roles are played by both individual isoforms and discrete protein domains.

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