scholarly journals Postmitotic expansion of cell nuclei requires nuclear actin filament bundling by α‐actinin 4

EMBO Reports ◽  
2020 ◽  
Vol 21 (11) ◽  
Author(s):  
Sylvia Krippner ◽  
Jannik Winkelmeier ◽  
Julian Knerr ◽  
Dominique T Brandt ◽  
David Virant ◽  
...  
2020 ◽  
Author(s):  
Sylvia Krippner ◽  
Jannik Winkelmeier ◽  
Carsten Schwan ◽  
Julian Knerr ◽  
David Virant ◽  
...  

AbstractThe actin cytoskeleton operates in a multitude of cellular processes including cell shape and migration, mechanoregulation, as well as membrane or organelle dynamics. However, its filamentous properties and functions inside the mammalian cell nucleus are less well explored. We previously described transient actin assembly at mitotic exit that promotes nuclear expansion during chromatin decondensation. Here, we identify non-muscle ACTN4 as a critical regulator to facilitate F-actin formation, reorganization and bundling during postmitotic nuclear expansion. ACTN4 binds to nuclear actin filaments and ACTN4 clusters associate with nuclear F-actin in a highly dynamic fashion. ACTN4 but not ACTN1 is required for proper postmitotic nuclear volume expansion, mediated by its actin binding domain. Using super-resolution imaging to quantify actin filament numbers and widths in individual nuclei we find that ACTN4 is necessary for postmitotic nuclear actin assembly and actin filament bundling. Our findings uncover a nuclear cytoskeletal function for ACTN4 to control nuclear size during mitotic cell division.


Author(s):  
Ivo Sauman

The non-histone proteins associated with eukaryotic nuclear chromatin are incompletely characterized and their function is poorly understood. Thirty years ago, the presence of actin in the eukaryotic nucleus was reported for first time. Since then, several biochemical studies have identified actin and myosin as significant constituents of isolated nuclear matrix from a variety of cells. The studies cited above and others make a strong case for presence of actin in nuclei, but do not implicate actin as a component of eukaryotic chromosomes.Our examination of cells associated with developing ovarian follicles of lepidopterans confirmed that under routine immunocytochemical conditions, no actin can be detected with anti-actin antibodies in the follicle cell nuclei (Fig. 1a) . However, Fig. 1b demonstrates that endonuclease pretreatment of the same preparation to remove DNA followed by anti-actin antibody staining uncovers the presence of nuclear actin. Moreover, by employing squash preparations of Drosophila salivary glands and the same endonuclease digestion, it is clear that the nuclear actin is directly associated with the polytene chromosomes (Fig. 2a,b).


1994 ◽  
Vol 125 (2) ◽  
pp. 369-380 ◽  
Author(s):  
K Cant ◽  
B A Knowles ◽  
M S Mooseker ◽  
L Cooley

Drosophila singed mutants were named for their gnarled bristle phenotype but severe alleles are also female sterile. Recently, singed protein was shown to have 35% peptide identity with echinoderm fascin. Fascin is found in actin filament bundles in microvilli of sea urchin eggs and in filopodial extensions in coelomocytes. We show that Drosophila singed is required for actin filament bundle formation in the cytoplasm of nurse cells during oogenesis; in severe mutants, the absence of cytoplasmic actin filament bundles allows nurse cell nuclei to lodge in ring canals and block nurse cell cytoplasm transport. Singed is also required for organized actin filament bundle formation in the cellular extension that forms a bristle; in severe mutants, the small disorganized actin filament bundles lack structural integrity and allow bristles to bend and branch during extension. Singed protein is also expressed in migratory cells of the developing egg chamber and in the socket cell of the developing bristle, but no defect is observed in these cells in singed mutants. Purified, bacterially expressed singed protein bundles actin filaments in vitro with the same stoichiometry reported for purified sea urchin fascin. Singed-saturated actin bundles have a molar ratio of singed/actin of approximately 1:4.3 and a transverse cross-banding pattern of 12 nm seen using electron microscopy. Our results suggest that singed protein is required for actin filament bundle formation and is a Drosophila homolog of echinoderm fascin.


2017 ◽  
Vol 1 (2) ◽  
pp. 155-158
Author(s):  
Samuela Cataldi ◽  
Andrea Lazzarini ◽  
Michela Codini ◽  
Giacomo Cascianelli ◽  
Alessandro Floridi ◽  
...  

Abstract Nuclear actin has been implicated in different nuclear functions. In this work, its localization in nuclear membrane, chromatin and nuclear lipid microdomains was investigated. The implication of sphingomyelin metabolism was studied. Nuclear membrane, chromatin and nuclear lipid microdomains were purified from hepatocyte nuclei and H35 human hepatoma cell nuclei. The presence of β-actin was analyzed with immunoblotting by using specific antibodies. Sphingomyelinase, sphingomyelin-synthase, and phosphatidylcholine-specific phospholipase C activities were assayed by using radioactivity sphingomyelin and phosphatidylcholine as substrate. The results showed that β-actin is localized in nuclear lipid microdomains and it increases in cancer cells. Evidence is provided to the difference of phosphatidylcholine and sphingomyelin metabolism in various subnuclear fractions of cancer cell nuclei compared with normal cells. Our findings show increase of sphingomyelin-synthase and inhibition of sphingomyelinase activity only in nuclear lipid microdomains. Nuclear lipid microdomains, constituted by phosphatidylcholine, sphingomyelin and cholesterol, play a role as platform for β-actin anchoring. Possible role of sphingomyelin metabolism in cancer cells is discussed.


2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Ying Wang ◽  
Alice Sherrard ◽  
Bing Zhao ◽  
Michael Melak ◽  
Jonathan Trautwein ◽  
...  

AbstractAlthough the properties of the actin cytoskeleton in the cytoplasm are well characterized, the regulation and function of nuclear actin filaments are only recently emerging. We previously demonstrated serum-induced, transient assembly of filamentous actin within somatic cell nuclei. However, the extracellular cues, cell surface receptors as well as underlying signaling mechanisms have been unclear. Here we demonstrate that physiological ligands for G protein-coupled receptors (GPCRs) promote nuclear F-actin assembly via heterotrimeric Gαq proteins. Signal-induced nuclear actin responses require calcium release from the endoplasmic reticulum (ER) targeting the ER-associated formin INF2 at the inner nuclear membrane (INM). Notably, calcium signaling promotes the polymerization of linear actin filaments emanating from the INM towards the nuclear interior. We show that GPCR and calcium elevations trigger nuclear actin-dependent alterations in chromatin organization, uncovering a general cellular mechanism by which physiological ligands and calcium promote nuclear F-actin assembly for rapid responses towards chromatin dynamics.


2019 ◽  
Vol 4 (31) ◽  
pp. eaav1987 ◽  
Author(s):  
N. Tsopoulidis ◽  
S. Kaw ◽  
V. Laketa ◽  
S. Kutscheidt ◽  
C. Baarlink ◽  
...  

T cell antigen receptor (TCR) signaling triggers selective cytokine expression to drive T cell proliferation and differentiation required for immune defense and surveillance. The nuclear signaling events responsible for specificity in cytokine gene expression upon T cell activation are largely unknown. Here, we uncover formation of a dynamic actin filament network in the nucleus that regulates cytokine expression for effector functions of CD4+T lymphocytes. TCR engagement triggers the rapid and transient formation of a nuclear actin filament network via nuclear Arp2/3 complex, induced by elevated nuclear Ca2+levels and regulated via N-Wasp and NIK. Specific interference with TCR-induced formation of nuclear actin filaments impairs production of effector cytokines and prevents generation of antigen-specific antibodies but does not interfere with immune synapse formation and cell proliferation. Ca2+-regulated actin polymerization in the nucleus allows CD4+T cells the rapid conversion of TCR signals into effector functions required for T cell help.


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