Lamin A Ser404 Is a Nuclear Target of Akt Phosphorylation in C2C12 Cells

Cenni, Vittoria; Bertacchini, Jessika; Beretti, Francesca; Lattanzi, Giovanna; Bavelloni, Alberto; Riccio, Massimo; Ruzzene, Maria; Marin, Oriano; Arrigoni, Giorgio; Parnaik, Veena K.; Wehnert, Manfred; Maraldi, N. M.; Pol, Anto De; Cocco, Lucio; Marmiroli, Sandra

doi:10.1021/pr800262g

Cited by 80 publications

(81 citation statements)

References 59 publications

(109 reference statements)

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“…AKT1 phosphorylates Lamin A/C at Serine 301 and 404, which targets it for degradation. 16,17 As we demonstrated previously, AKT1 is expressed during late terminal differentiation in the mouse, 14 and this correlates with increased expression of epidermal terminal differentiation markers between E15.5 and E18.5 of mouse embryonic development. 14 Therefore, we investigated whether nuclear size change and the phosphorylation of Lamin A/C also occurs during this time.…”

supporting

confidence: 65%

“…AKT1 kd plasmids were transfected into rat epidermal keratinocyte (REK) cells, using lipofectamine (Invitrogen), according to the manufacturer's instructions. The S404A and S301A Lamin A constructs have been previously described 16,17 and transfected into REK cells in an identical fashion. Cells were cultured and selected for two weeks in 100 μM G418 (Gibco).…”

Section: Discussionmentioning

confidence: 99%

“…Antibodies used were rabbit anti-pSer 473 AKT (1 : 10, Cell Signalling), mouse anti-AKT1 (1 : 10; Cell Signalling), mouse anti-Flag (1 : 10, Cell Signaling Technologies), rabbit antifilaggrin (1 : 50, Santa Cruz Biotechnologies), rabbit anti-Loricrin (1 : 100; Covance), mouse anti-Lamin A/C (1 : 200, Santa Cruz Biotechnologies), and rabbit anti-SMAD 1 (1 : 25 Epitomics). The rabbit anti-lamin A phospho-serine 404 antibody was described previously 16,17 ) and used at 1 : 25. Primary antibodies were detected using Alexa 488 and 594-conjugated goat anti-mouse and anti-rabbit (Invitrogen, 1 : 500).…”

Section: Discussionmentioning

confidence: 99%

“…Lamin A is a known target of AKT, and AKT phosphorylation is required for Lamin A degradation in other tissues. 16,17 To test the consequences of expressing a non-degradable Lamin A on keratinocyte differentiation, we transfected cells with human flag-tagged Lamin A with serine to alanine mutations at positions 301 and 404 (S301A and S404A; Figure 6a). Both prevented degradation of Lamin A, which was shown by the immuno-depletion experiments to be construct specific; that is, the mutant Lamin A did not inhibit or prevent the degradation of the wild-type Lamin A/C present ( Figure 6a).…”

mentioning

confidence: 99%

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AKT1-mediated Lamin A/C degradation is required for nuclear degradation and normal epidermal terminal differentiation

Naeem¹,

Zhu²,

Di³

et al. 2015

Cell Death Differ

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Nuclear degradation is a key stage in keratinocyte terminal differentiation and the formation of the cornified envelope that comprises the majority of epidermal barrier function. Parakeratosis, the retention of nuclear material in the cornified layer of the epidermis, is a common histological observation in many skin diseases, notably in atopic dermatitis and psoriasis. Keratinocyte nuclear degradation is not well characterised, and it is unclear whether the retained nuclei contribute to the altered epidermal differentiation seen in eczema and psoriasis. Loss of AKT1 function strongly correlated with parakeratosis both in eczema samples and in organotypic culture models. Although levels of DNAses, including DNase1L2, were unchanged, proteomic analysis revealed an increase in Lamin A/C. AKT phosphorylates Lamin A/C, targeting it for degradation. Consistent with this, Lamin A/C degradation was inhibited and Lamin A/C was observed in the cornified layer of AKT1 knockdown organotypic cultures, surrounding retained nuclear material. Using AKT-phosphorylation-dead Lamin A constructs we show that the retention of nuclear material is sufficient to cause profound changes in epidermal terminal differentiation, specifically a reduction in Loricrin, Keratin 1, Keratin 10, and filaggrin expression. We show that preventing nuclear degradation upregulates BMP2 expression and SMAD1 signalling. Consistent with these data, we observe both parakeratosis and evidence of increased SMAD1 signalling in atopic dermatitis. We therefore present a model that, in the absence of AKT1-mediated Lamin A/C degradation, DNA degradation processes, such as those mediated by DNAse 1L2, are prevented, leading to parakeratosis and changes in epidermal differentiation. Nuclear degradation is a key stage in keratinocyte terminal differentiation and the formation of the cornified envelope that comprises the majority of epidermal barrier function. [1][2][3] Parakeratosis, the retention of nuclear material in the cornified layer of the epidermis, is a common histological observation in many skin diseases, but most notably in the epidermal barrierdefective diseases eczema and psoriasis. 4,5 Mechanisms of nuclear degradation in the epidermis have not yet been well characterised and it is not known whether the retained nuclei contribute to the altered epidermal differentiation programmes seen in these skin diseases. 6,7 It is surprising that, for such a critical component of epidermal terminal differentiation, relatively few molecular mechanisms inducing parakeratosis have been investigated. The caspase-14 knockout mouse develops parakeratotic plaques upon chemical barrier disruption 8 and has subtle defects in epidermal terminal differentiation, including filaggrin processing, 9 whereas the DNAse 1L2 knockout mouse showed constitutive nuclear retention in hair and nails, which led to structural abnormalities in the hair shaft. 10,11 Parakeratosis also occurs during wound healing. 12 Nuclei are retained in the scab of healing wounds, and this correlates with...

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supporting

confidence: 65%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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AKT1-mediated Lamin A/C degradation is required for nuclear degradation and normal epidermal terminal differentiation

Naeem¹,

Zhu²,

Di³

et al. 2015

Cell Death Differ

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“…For instance, lamin A has been shown to be phosphorylated at its S404 residue by Akt/PKB kinase downstream of the phosphoinositide-3-kinase (PI3 kinase) signaling pathway, in response to insulin stimulation in myoblasts. In cells from an EDMD-2 patient carrying a mutation at Arg401, which lies at the Akt consensus, lamin phosphorylation was dramatically reduced [124]. The pathological mechanism of such an effect is unknown.…”

Section: Tissue Selective Phenotyphesmentioning

confidence: 99%

Laminopathies: The molecular background of the disease and the prospects for its treatment

Zaremba-Czogalla

Dubińska–Magiera

Rzepecki

2011

Cellular and Molecular Biology Letters

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phone: +48 71 375 63 08Abbreviations used: ADLD -adult-onset autosomal dominant leukodystrophy; APLacquired partial lipodystrophy; CMD1A -dilated cardiomyopathy 1A with conduction defect; CMT2B1 Charcot-Marie-Tooth disease type 2B1; DMD -Duchenne muscular dystrophy; EDMD -autosomal dominant Emery-Dreifuss muscular dystrophy; ERKextracellular signal-regulated kinase; GL -generalized lipodystrophy; FPLD -Dunnigan familial partial lipodystrophy; HGPS -Hutchinson Gilford progeria syndrome; INMinner nuclear membrane; JNK -c-Jun NH(2)-terminal kinase; LAP -lamina-associated polypeptide; LBR -p58 protein, lamin B receptor, 3 beta-hydroxysterol D14-reductase;LGMDB1 -limb-girdle muscular dystrophy type 1B; LINC -linker of the nucleoskeleton and cytoskeleton; LMNA -gene encoding A/C type lamins; MAD -mandibuloacral dysplasia; NE -nuclear envelope; PPARγ -peroxisome proliferator-activated receptor γ; pRB -retinoblastoma protein; ZMPSTE24/FACE1 -zinc metalloproteinase STE 24 homology Review LAMINOPATHIES: THE MOLECULAR BACKGROUND OF THE DISEASE AND THE PROSPECTS FOR ITS TREATMENTMAGDALENA ZAREMBA-CZOGALLA, MAGDA DUBIŃSKA-MAGIERA and RYSZARD RZEPECKI* Laboratory of Nuclear Proteins, Faculty of Biotechnology, University of Wrocław, ul. Przybyszewskiego 63/77, 51-148 Wroclaw, PolandAbstract: Laminopathies are rare human degenerative disorders with a wide spectrum of clinical phenotypes, associated with defects in the main protein components of the nuclear envelope, mostly in the lamins. They include systemic disorders and tissue-restricted diseases. Scientists have been trying to explain the pathogenesis of laminopathies and find an efficient method for treatment for many years. In this review, we discuss the current state of knowledge about laminopathies, the molecular mechanisms behind the development of particular phenotypes, and the prospects for stem cell and/or gene therapy treatments.

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Amniotic fluid stem cell exosomes: Therapeutic perspective

et al. 2018

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It is widely accepted that the therapeutic potential of stem cells can be largely mediated by paracrine factors, also included into exosomes. Thus, stem cell-derived exosomes represent a major therapeutic option in regenerative medicine avoiding, if compared to stem cells graft, abnormal differentiation and tumor formation. Exosomes derived from mesenchymal stem cells (MSC) induce damaged tissue repair, and can also exert immunomodulatory effects on the differentiation, activation and function of different lymphocytes. Therefore, MSC exosomes can be considered as a potential treatment for inflammatory diseases and also an ideal candidate for allogeneic therapy due to their low immunogenicity. Amniotic fluid stem cells (AFSCs) are broadly multipotent, can be expanded in culture, and can be easily cryopreserved in cellular banks. In this study, morphology, phenotype, and protein content of exosomes released into amniotic fluid in vivo and from AFSC during in vitro culture (conditioned medium) were examined. We found that AFSC-derived exosomes present different molecules than amniotic fluid ones, some of them involved in immunomodulation, such transforming growth factor beta and hepatic growth factors. The immunomodulatory effect of AFSC's exosomes on peripheral blood mononuclear cells stimulated with phytohemagglutinin was compared to that of the supernatant produced by such conditioned media deprived of exosomes. We present evidence that the principal effect of AFSC conditioned media (without exosomes) is the induction of apoptosis in lymphocytes, whereas exposure to AFSC-derived exosomes decreases the lymphocyte's proliferation, supporting the hypothesis that the entire secretome of stem cells differently affects immune-response. © 2017 BioFactors, 44(2):158-167, 2018.

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Lamin A Ser404 Is a Nuclear Target of Akt Phosphorylation in C2C12 Cells

Cited by 80 publications

References 59 publications

AKT1-mediated Lamin A/C degradation is required for nuclear degradation and normal epidermal terminal differentiation

AKT1-mediated Lamin A/C degradation is required for nuclear degradation and normal epidermal terminal differentiation

Laminopathies: The molecular background of the disease and the prospects for its treatment

Amniotic fluid stem cell exosomes: Therapeutic perspective

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