scholarly journals Phosphorylation and dephosphorylation of Ser852 and Ser889 control the clustering, localization and function of PAR3

2020 ◽  
Vol 133 (22) ◽  
pp. jcs244830
Author(s):  
Kazunari Yamashita ◽  
Keiko Mizuno ◽  
Kana Furukawa ◽  
Hiroko Hirose ◽  
Natsuki Sakurai ◽  
...  
Keyword(s):  
Tight Junction ◽  
Protein Phosphatase ◽  
Phosphorylation Site ◽  
Protein Phosphatase 1 ◽  
Membrane Domain ◽  
Unique Role ◽  
Cellular Events ◽  
Local Clustering ◽  
And Function ◽  
Specific Membrane

ABSTRACTCell polarity is essential for various asymmetric cellular events, and the partitioning defective (PAR) protein PAR3 (encoded by PARD3 in mammals) plays a unique role as a cellular landmark to establish polarity. In epithelial cells, PAR3 localizes at the subapical border, such as the tight junction in vertebrates, and functions as an apical determinant. Although we know a great deal about the regulators of PAR3 localization, how PAR3 is concentrated and localized to a specific membrane domain remains an important question to be clarified. In this study, we demonstrate that ASPP2 (also known as TP53BP2), which controls PAR3 localization, links PAR3 and protein phosphatase 1 (PP1). The ASPP2–PP1 complex dephosphorylates a novel phosphorylation site, Ser852, of PAR3. Furthermore, Ser852- or Ser889-unphosphorylatable PAR3 mutants form protein clusters, and ectopically localize to the lateral membrane. Concomitance of clustering and ectopic localization suggests that PAR3 localization is a consequence of local clustering. We also demonstrate that unphosphorylatable forms of PAR3 exhibited a low molecular turnover and failed to coordinate rapid reconstruction of the tight junction, supporting that both the phosphorylated and dephosphorylated states are essential for the functional integrity of PAR3.

scholarly journals Phosphorylation and dephosphorylation of Ser852 and Ser889 control clustering, localization, and function of PAR-3

2020 ◽  
Author(s):  
Kazunari Yamashita ◽  
Keiko Mizuno ◽  
Kana Furukawa ◽  
Hiroko Hirose ◽  
Natsuki Sakurai ◽  
...  
Keyword(s):  
Tight Junction ◽  
Cell Polarity ◽  
Phosphorylation Site ◽  
Cell Junction ◽  
Unique Role ◽  
Cellular Events ◽  
A Cell ◽  
Summary Statement ◽  
And Function ◽  
Specific Membrane

AbstractCell polarity is essential for various asymmetric cellular events, where the partitioning defective (PAR) protein, PAR3, plays a unique role as a cellular landmark to establish polarity. In epithelial cells, PAR3 localizes at the subapical border such as the tight junction in vertebrates and functions as an apical determinant. Although there is much information about the regulators of PAR3 localization, the mechanism involved in PAR3 concentration and localization to the specific membrane domain remains an important question to be clarified. In this study, we demonstrate that ASPP2, a stimulator of PAR3 localization, can link PAR3 and protein phosphatase 1 (PP1). The ASPP2–PP1 complex dephosphorylates a novel phosphorylation site, Ser852, of PAR3. Furthermore, Ser852- or Ser889-unphosphorylatable PAR3 mutants form protein clusters and ectopically localize to the lateral membrane. Concomitance of clustering and ectopic localization suggests that PAR3 localization is a consequence of local clustering. We also demonstrate that unphosphorylatable forms of PAR3 are static in molecular turnover and fail to coordinate rapid reconstruction of the tight junction, supporting that both phosphorylated and dephosphorylated states are essential for the functional integrity of PAR3.Summary statementWe show that phosphorylation and dephosphorylation regulate clustering of PAR-3, a cell polarity-regulating factor, and how the clustering regulation affects localization of PAR-3 and cell-cell junction formation.

scholarly journals Protein Phosphatase 1 Inhibitor–1 Mediates the cAMP-Dependent Stimulation of the Renal NaCl Cotransporter

2019 ◽  
Vol 30 (5) ◽  
pp. 737-750 ◽  
Author(s):  
David Penton ◽  
Sandra Moser ◽  
Agnieszka Wengi ◽  
Jan Czogalla ◽  
Lena Lindtoft Rosenbaek ◽  
...  
Keyword(s):  
Protein Phosphatase ◽  
Mdck Cells ◽  
Ex Vivo ◽  
Phosphorylation Site ◽  
Mouse Kidney ◽  
Protein Phosphatase 1 ◽  
Endocrine Disorders ◽  
Kidney Slices ◽  
Nacl Cotransporter ◽  
Stimulation Of

BackgroundA number of cAMP-elevating hormones stimulate phosphorylation (and hence activity) of the NaCl cotransporter (NCC) in the distal convoluted tubule (DCT). Evidence suggests that protein phosphatase 1 (PP1) and other protein phosphatases modulate NCC phosphorylation, but little is known about PP1’s role and the mechanism regulating its function in the DCT.MethodsWe used ex vivo mouse kidney preparations to test whether a DCT-enriched inhibitor of PP1, protein phosphatase 1 inhibitor–1 (I1), mediates cAMP’s effects on NCC, and conducted yeast two-hybrid and coimmunoprecipitation experiments in NCC-expressing MDCK cells to explore protein interactions.ResultsTreating isolated DCTs with forskolin and IBMX increased NCC phosphorylation via a protein kinase A (PKA)–dependent pathway. Ex vivo incubation of mouse kidney slices with isoproterenol, norepinephrine, and parathyroid hormone similarly increased NCC phosphorylation. The cAMP-induced stimulation of NCC phosphorylation strongly correlated with the phosphorylation of I1 at its PKA consensus phosphorylation site (a threonine residue in position 35). We also found an interaction between NCC and the I1-target PP1. Moreover, PP1 dephosphorylated NCC in vitro, and the PP1 inhibitor calyculin A increased NCC phosphorylation. Studies in kidney slices and isolated perfused kidneys of control and I1-KO mice demonstrated that I1 participates in the cAMP-induced stimulation of NCC.ConclusionsOur data suggest a complete signal transduction pathway by which cAMP increases NCC phosphorylation via a PKA-dependent phosphorylation of I1 and subsequent inhibition of PP1. This pathway might be relevant for the physiologic regulation of renal sodium handling by cAMP-elevating hormones, and may contribute to salt-sensitive hypertension in patients with endocrine disorders or sympathetic hyperactivity.

Identification and characterization of cAMP-dependent protein kinase and its possible direct interactions with protein phosphatase-1 in marine dinoflagellates

1996 ◽  
Vol 74 (4) ◽  
pp. 559-567 ◽  
Author(s):  
John F. Dawson ◽  
Kathy He Wang ◽  
Charles F. B. Holmes
Keyword(s):  
Signal Transduction ◽  
Protein Kinase ◽  
Okadaic Acid ◽  
Protein Phosphatase ◽  
Phosphorylation Site ◽  
Protein Phosphatase 1 ◽  
Dependent Protein Kinase ◽  
Prorocentrum Lima ◽  
Camp Dependent Protein Kinase ◽  
Dependent Protein

We have examined the nature of signal transduction involving reversible protein phosphorylation in marine Prorocentrale species. Of particular interest is the marine dinoflagellate Prorocentrum lima in which the tumour promoter okadaic acid is produced and may interfere with signal transduction. We have identified cAMP-dependent protein kinase (PKA) activity in P. lima, P. micans, and P. minimum. The P. lima enzyme was characterized biochemically and appears to consist of two different isoforms in the R2C2 configuration. Whole cell extracts of P. micans and P. minimum treated with the specific PKA inhibitor peptide PKI (5–24) or cAMP demonstrated altered intensities of phosphopeptide 32P labeling, most likely involving regulation of a protein phosphatase via PKA activity. A primary candidate for PKA regulation is protein phosphatase-1 (PP-1), which in P. lima possesses a classical PKA consensus phosphorylation site. We demonstrate that a peptide fragment of PP-1 from P. lima corresponding to this PKA phosphorylation site can be effectively phosphorylated by PKA and dephosphorylated by calcineurin. We speculate that PP-1 activity among several lower eukaryotes may be mediated directly by reversible phosphorylation. Higher eukaryotes may have developed inhibitor proteins to provide more complex regulation of protein phosphatase activity.Key words: cAMP-dependent protein kinase, protein phosphatase-1, dinoflagellates, Prorocentrum lima, okadaic acid.

scholarly journals Molecular Investigations of the Structure and Function of the Protein Phosphatase 1−Spinophilin−Inhibitor 2 Heterotrimeric Complex

Biochemistry ◽  
2011 ◽  
Vol 50 (7) ◽  
pp. 1238-1246 ◽  
Author(s):  
Barbara Dancheck ◽  
Michael J. Ragusa ◽  
Marc Allaire ◽  
Angus C. Nairn ◽  
Rebecca Page ◽  
...  
Keyword(s):  
Protein Phosphatase ◽  
Structure And Function ◽  
Protein Phosphatase 1 ◽  
And Function

scholarly journals Protein phosphatase 1 regulates assembly and function of the β-catenin degradation complex

2007 ◽  
Vol 26 (6) ◽  
pp. 1511-1521 ◽  
Author(s):  
Wen Luo ◽  
Annita Peterson ◽  
Benjamin A Garcia ◽  
Gary Coombs ◽  
Bente Kofahl ◽  
...  
Keyword(s):  
Protein Phosphatase ◽  
Protein Phosphatase 1 ◽  
And Function

scholarly journals Degeneracy and Function of the Ubiquitous RVXF Motif That Mediates Binding to Protein Phosphatase-1

2003 ◽  
Vol 278 (21) ◽  
pp. 18817-18823 ◽  
Author(s):  
Paulina Wakula ◽  
Monique Beullens ◽  
Hugo Ceulemans ◽  
Willy Stalmans ◽  
Mathieu Bollen
Keyword(s):  
Protein Phosphatase ◽  
Protein Phosphatase 1 ◽  
And Function ◽  
Rvxf Motif

scholarly journals The Oncogenic Potential of TCTEX1D4 is Modulated by the Phosphoprotein Phosphatase PP1

2021 ◽  
Author(s):  
Juliana Felgueiras ◽  
Luís Sousa ◽  
Ana Luísa Luísa Teixeira ◽  
Bárbara Regadas ◽  
Luís Korrodi-Gregório ◽  
...  
Keyword(s):  
Protein Phosphatase ◽  
Protein Phosphatase 1 ◽  
Human Prostate ◽  
Interacting Proteins ◽  
Interacting Protein ◽  
Oncogenic Potential ◽  
Regulatory Subunits ◽  
Prostate Tumorigenesis ◽  
Cellular Events

Abstract Protein phosphatase 1 (PP1) regulates several cellular events via interaction with multiple regulatory subunits. The human prostate proteome includes various PP1-interacting proteins; however, a very limited number of interactions is yet characterized and their role in prostate tumorigenesis remains poorly understood. Tctex1 domain-containing protein 4 (TCTEX1D4) was previously identified as a PP1-interacting protein, but its function, as well as the relevance of its interaction with PP1, are virtually unknown. In this study we addressed the role of the PP1/TCTEX1D4 complex in prostate tumorigenesis. We found distinct expression levels and subcellular distributions for TCTEX1D4 and PP1γ in human prostate epithelial normal-like and malignant cells. Moreover, we showed that TCTEX1D4 participates in the regulation of cell proliferation and modulation of microRNAs expression and that its interaction with PP1 controls its function. Taken together, our study provides first evidence for the involvement of the PP1/TCTEX1D4 complex in prostate tumorigenesis.

Biogenesis and activity regulation of protein phosphatase 1

2017 ◽  
Vol 45 (1) ◽  
pp. 89-99 ◽  
Author(s):  
Iris Verbinnen ◽  
Monica Ferreira ◽  
Mathieu Bollen
Keyword(s):  
Protein Phosphatase ◽  
Protein Phosphatase 1 ◽  
Interacting Proteins ◽  
Subcellular Targeting ◽  
Substantial Fraction ◽  
Short Linear Motifs ◽  
Specific Subset ◽  
Linear Motifs ◽  
And Function ◽  
And Control

Protein phosphatase 1 (PP1) is expressed in all eukaryotic cells and catalyzes a substantial fraction of phosphoserine/threonine dephosphorylation reactions. It forms stable complexes with PP1-interacting proteins (PIPs) that guide the phosphatase throughout its life cycle and control its fate and function. The diversity of PIPs is huge (≈200 in vertebrates), and most of them combine short linear motifs to form large and unique interaction interfaces with PP1. Many PIPs have separate domains for PP1 anchoring, PP1 regulation, substrate recruitment and subcellular targeting, which enable them to direct associated PP1 to a specific subset of substrates and mediate acute activity control. Hence, PP1 functions as the catalytic subunit of a large number of multimeric holoenzymes, each with its own subset of substrates and mechanism(s) of regulation.

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