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Phactr1/PP1 phosphatase substrates in mouse primary neurons

ABSTRACT: Neurons express Phactr1 at high level (Allen et al., 2004), and Phactr1 mutations are associated with morphological and functional developmental defects in cortical neurons (Hamada et al., 2018). To identify targets of the Phactr1/PP1 phosphatase holoenzyme, we cultured hippocampal and cortical neurons from wildtype and Phactr1-null animals, treated them either with Latrunculin B or Cytochalasin D to inhibit or activate formation of the Phactr1/PP1 complex (Wiezlak et al., 2012), and analysed phosphorylation profiles using TMT phosphoproteomics. Among over 9000 phosphorylation sites quantified, we found numerous phosphorylation sites that differed significantly in their response to these stimuli in wildtype as opposed to Phactr1-null neurons.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Brain, Neuron

SUBMITTER: Roman Fedoryshchak

LAB HEAD: Richard Treisman

PROVIDER: PXD019882 | Pride | 2020-10-28

REPOSITORIES: Pride

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Molecular basis for substrate specificity of the Phactr1/PP1 phosphatase holoenzyme.

Fedoryshchak Roman O RO Přechová Magdalena M Butler Abbey M AM Lee Rebecca R O'Reilly Nicola N Flynn Helen R HR Snijders Ambrosius P AP Eder Noreen N Ultanir Sila S Mouilleron Stephane S Treisman Richard R

eLife 20200925

PPP-family phosphatases such as PP1 have little intrinsic specificity. Cofactors can target PP1 to substrates or subcellular locations, but it remains unclear how they might confer sequence-specificity on PP1. The cytoskeletal regulator Phactr1 is a neuronally enriched PP1 cofactor that is controlled by G-actin. Structural analysis showed that Phactr1 binding remodels PP1's hydrophobic groove, creating a new composite surface adjacent to the catalytic site. Using phosphoproteomics, we identified ...[more]

PMID: 32975518

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