Mapping the contact surfaces in the Lamin A:AIMP3 complex by hydrogen/deuterium exchange FT-ICR mass spectrometry

Tao, Yeqing; Fang, Pengfei; Kim, Sung Hoon; Guo, Min; Young, Nicolas L.; Marshall, Alan G.

doi:10.1371/journal.pone.0181869

Cited by 7 publications

(10 citation statements)

References 49 publications

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“…Our results indicate that the Eef1e1 is another TCR signal strength-responsive driver of the Th1 cell fate. Eef1e1 binds to Lamin A (61), and overexpression studies performed in cell lines demonstrated that Eef1e1 facilitates Lamin A ubiquitination and degradation via the ubiquitin ligase, Siah1 (62). Interestingly, Lamin A-deficient T cells have reduced in vivo Th1 cell differentiation (63), whereas Siah1 promotes Th17 cell differentiation in vitro (64).…”

Section: Discussionmentioning

confidence: 99%

TCR Affinity Biases Th Cell Differentiation by Regulating CD25, Eef1e1, and Gbp2

Kotov

Mitchell

Pengo

et al. 2019

The Journal of Immunology

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Naive CD4 + T lymphocytes differentiate into various Th cell subsets following TCR binding to microbial peptide:MHC class II (p:MHCII) complexes on dendritic cells (DCs). The affinity of the TCR interaction with p:MHCII plays a role in Th differentiation by mechanisms that are not completely understood. We found that low-affinity TCRs biased mouse naive T cells to become T follicular helper (Tfh) cells, whereas higher-affinity TCRs promoted the formation of Th1 or Th17 cells. We explored the basis for this phenomenon by focusing on IL-2R signaling, which is known to promote Th1 and suppress Tfh cell differentiation. SIRP⍺ + DCs produce abundant p:MHCII complexes and consume IL-2, whereas XCR1 + DCs weakly produce p:MHCII but do not consume IL-2. We found no evidence, however, of preferential interactions between Th1 cell-prone, high-affinity T cells and XCR1 + DCs or Tfh cell-prone, low-affinity T cells and SIRP⍺ + DCs postinfection with bacteria expressing the peptide of interest. Rather, high-affinity T cells sustained IL-2R expression longer and expressed two novel Th cell differentiation regulators, Eef1e1 and Gbp2, to a higher level than low-affinity T cells. These results suggest that TCR affinity does not influence Th cell differentiation by biasing T cell interactions with IL-2-consuming DCs, but instead, directly regulates genes in naive T cells that control the differentiation process.

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Section: Discussionmentioning

confidence: 99%

TCR Affinity Biases Th Cell Differentiation by Regulating CD25, Eef1e1, and Gbp2

Kotov

Mitchell

Pengo

et al. 2019

The Journal of Immunology

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“…These partners include LAP1β (disrupted by lamin A variant p.G602S), ZNF3 (disrupted by p.G602S, p.G608S, p.T623S, p.R644C), MORF4L1 (disrupted by p.G602S, p.G608S, p.T623S), Cyclin G1 (disrupted by p.G602S, p.G608S, p.R644C), and CENP-P (disrupted by p.R644C) [ 90 ]. We speculate that ‘sweet-spot’ modifications might also influence association with AIMP3, a cytoplasmic scaffolding protein that can also enter the nucleus and act as a tumor suppressor [ 91 , 92 ]. AIMP3 binds directly to residues 641–647 in the ‘sweet spot’ of mature lamin A [ 91 ].…”

Section: Discussionmentioning

confidence: 99%

“…We speculate that ‘sweet-spot’ modifications might also influence association with AIMP3, a cytoplasmic scaffolding protein that can also enter the nucleus and act as a tumor suppressor [ 91 , 92 ]. AIMP3 binds directly to residues 641–647 in the ‘sweet spot’ of mature lamin A [ 91 ]. AIMP3 recruits Siah1 (an E3 ubiquitin ligase) and triggers selective degradation of mature lamin A to promote senescence [ 91 , 92 ].…”

Section: Discussionmentioning

confidence: 99%

OGT (O-GlcNAc Transferase) Selectively Modifies Multiple Residues Unique to Lamin A

Simon

Wriston

Fan

et al. 2018

Cells

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The LMNA gene encodes lamins A and C with key roles in nuclear structure, signaling, gene regulation, and genome integrity. Mutations in LMNA cause over 12 diseases (‘laminopathies’). Lamins A and C are identical for their first 566 residues. However, they form separate filaments in vivo, with apparently distinct roles. We report that lamin A is β-O-linked N-acetylglucosamine-(O-GlcNAc)-modified in human hepatoma (Huh7) cells and in mouse liver. In vitro assays with purified O-GlcNAc transferase (OGT) enzyme showed robust O-GlcNAcylation of recombinant mature lamin A tails (residues 385–646), with no detectable modification of lamin B1, lamin C, or ‘progerin’ (Δ50) tails. Using mass spectrometry, we identified 11 O-GlcNAc sites in a ‘sweet spot’ unique to lamin A, with up to seven sugars per peptide. Most sites were unpredicted by current algorithms. Double-mutant (S612A/T643A) lamin A tails were still robustly O-GlcNAc-modified at seven sites. By contrast, O-GlcNAcylation was undetectable on tails bearing deletion Δ50, which causes Hutchinson–Gilford progeria syndrome, and greatly reduced by deletion Δ35. We conclude that residues deleted in progeria are required for substrate recognition and/or modification by OGT in vitro. Interestingly, deletion Δ35, which does not remove the majority of identified O-GlcNAc sites, does remove potential OGT-association motifs (lamin A residues 622–625 and 639–645) homologous to that in mouse Tet1. These biochemical results are significant because they identify a novel molecular pathway that may profoundly influence lamin A function. The hypothesis that lamin A is selectively regulated by OGT warrants future testing in vivo, along with two predictions: genetic variants may contribute to disease by perturbing OGT-dependent regulation, and nutrient or other stresses might cause OGT to misregulate wildtype lamin A.

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“…Normally found in the cytoplasm with roles in translation, AIMP3 also enters the nucleus and acts as a tumor suppressor (79). AIMP3 binds directly to residues 641-647 in the 'sweet spot' of mature lamin A (78). AIMP3 recruits Siah1 (an E3 ubiquitin ligase) and triggers selective degradation of mature lamin A to promote senescence (78,79).…”

Section: Potential Impacts Of O-glcnac On Mature Lamin a Functionmentioning

confidence: 99%

“…AIMP3 binds directly to residues 641-647 in the 'sweet spot' of mature lamin A (78). AIMP3 recruits Siah1 (an E3 ubiquitin ligase) and triggers selective degradation of mature lamin A to promote senescence (78,79).…”

Section: Potential Impacts Of O-glcnac On Mature Lamin a Functionmentioning

confidence: 99%

OGT (O-GlcNAc Transferase) selectively modifies multiple residues unique to lamin A

Simon

Wriston

Fan

et al. 2017

Preprint

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The LMNA gene encodes lamins A and C with key roles in nuclear structure, signaling, chromatin organization, and genome integrity. Mutations in LMNA cause >12 diseases, termed laminopathies.Lamins A and C are identical for their first 566 residues. However, they form distinct filaments in vivo with apparently distinct roles. We report that lamin A is O-GlcNAc modified in human hepatoma (Huh7) cells and in mouse liver. In vitro assays with purified OGT enzyme showed robust O-GlcNAcylation of recombinant mature lamin A tails (residues 385-646), with no detectable modification of lamin B1, lamin C, or 'progerin' (Δ50) tails. Using mass spectrometry, we identified 11 O-GlcNAc sites in a 'sweet spot' unique to lamin A, with up to seven sugars per peptide. Most sites were unpredicted by current algorithms. Double-mutant (S612A/T643A) lamin A tails were still robustly O-GlcNAc-modified at seven sites. By contrast, O-GlcNAcylation was undetectable on tails bearing deletion Δ50, which causes Hutchinson-Gilford progeria syndrome, and greatly reduced by deletion Δ35, suggesting this region is required for substrate recognition or modification by OGT in vitro. These results suggest OGT, an essential protein and master regulator, regulates partners or function(s) unique to lamin A that are lost in progeria.

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Mapping the contact surfaces in the Lamin A:AIMP3 complex by hydrogen/deuterium exchange FT-ICR mass spectrometry

Cited by 7 publications

References 49 publications

TCR Affinity Biases Th Cell Differentiation by Regulating CD25, Eef1e1, and Gbp2

TCR Affinity Biases Th Cell Differentiation by Regulating CD25, Eef1e1, and Gbp2

OGT (O-GlcNAc Transferase) Selectively Modifies Multiple Residues Unique to Lamin A

OGT (O-GlcNAc Transferase) selectively modifies multiple residues unique to lamin A

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