scholarly journals Keratins and plakin family cytolinker proteins control the length of epithelial microridge protrusions

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Yasuko Inaba ◽  
Vasudha Chauhan ◽  
Aaron Paul van Loon ◽  
Lamia Saiyara Choudhury ◽  
Alvaro Sagasti
Keyword(s):  
Intermediate Filaments ◽  
Intermediate Filament ◽  
Actin Filaments ◽  
Actin Binding ◽  
Protein Levels ◽  
Skin Cells ◽  
Keratin Filaments ◽  
Actin Binding Domain ◽  
Cytoskeletal Elements ◽  
Keratin Intermediate Filaments

Actin filaments and microtubules create diverse cellular protrusions, but intermediate filaments, the strongest and most stable cytoskeletal elements, are not known to directly participate in the formation of protrusions. Here we show that keratin intermediate filaments directly regulate the morphogenesis of microridges, elongated protrusions arranged in elaborate maze-like patterns on the surface of mucosal epithelial cells. We found that microridges on zebrafish skin cells contained both actin and keratin filaments. Keratin filaments stabilized microridges, and overexpressing keratins lengthened them. Envoplakin and periplakin, plakin family cytolinkers that bind F-actin and keratins, localized to microridges, and were required for their morphogenesis. Strikingly, plakin protein levels directly dictate microridge length. An actin-binding domain of periplakin was required to initiate microridge morphogenesis, whereas periplakin-keratin binding was required to elongate microridges. These findings separate microridge morphogenesis into distinct steps, expand our understanding of intermediate filament functions, and identify microridges as protrusions that integrate actin and intermediate filaments.

scholarly journals Keratins and Plakin family cytolinker proteins control the length of epithelial microridge protrusions

2020 ◽  
Author(s):  
Yasuko Inaba ◽  
Vasudha Chauhan ◽  
Aaron Paul van Loon ◽  
Lamia Saiyara Choudhury ◽  
Alvaro Sagasti
Keyword(s):  
Intermediate Filaments ◽  
Actin Filaments ◽  
Actin Binding ◽  
Protein Levels ◽  
Skin Cells ◽  
Keratin Filaments ◽  
Actin Binding Domain ◽  
Stable Class ◽  
Cytoskeletal Elements ◽  
Keratin Intermediate Filaments

ABSTRACTActin filaments and microtubules create diverse cellular protrusions, but intermediate filaments, the strongest and most stable class of cytoskeletal elements, are not known to directly participate in the formation of protrusions. Here we show that Keratin intermediate filaments directly regulate the morphogenesis of microridges, elongated protrusions from mucosal epithelial cells arranged in elaborate fingerprint-like patterns. Developing microridges on zebrafish skin cells contained both Actin and Keratin filaments. Keratin filaments maintained microridges upon F-actin disruption, and overexpressing Keratins lengthened microridges. Envoplakin and Periplakin, Plakin family cytolinkers that bind to F-actin and Keratins, localized to microridges and were required for their morphogenesis. Strikingly, Plakin protein levels directly determined microridge length. An actin-binding domain of Periplakin was required to initiate microridge morphogenesis, whereas Periplakin-Keratin binding was required to stabilize and elongate microridges. Our results thus separate microridge morphogenesis into two steps with differential requirements for cytoskeletal elements, expand our understanding of intermediate filament functions, and identify microridges as cellular protrusions that integrate actin and intermediate filaments.

Actin Filaments Decorated with Cytoskeletal Proteins

1998 ◽  
Vol 4 (S2) ◽  
pp. 458-459
Author(s):  
D. Hanein ◽  
S. Goldsmith ◽  
W. Lehman ◽  
R. Craig ◽  
I. Correia ◽  
...  
Keyword(s):  
Intermediate Filaments ◽  
Intermediate Filament ◽  
Actin Filaments ◽  
Three Dimensional ◽  
Cytoskeletal Proteins ◽  
Actin Binding ◽  
Structural Systems ◽  
Calponin Homology Domain ◽  
Flexible Molecules ◽  
Actin Binding Domain

A superfamily of actin crosslinking proteins, the calponin-homology domain (CH) family is implicated in directing the assembly and controlling the organization of the cytoskeleton through a highly conserved actin binding domain (ABD). This domain, composed of a tandem pair of CH motifs, is found in actin crosslinking proteins such as fimbrin, alpha-actinin, spectrin, and dystrophin. In addition to its role in crosslinking actin filaments, a putative homologous ABD in intermediate filament-binding proteins, plectin, dystonin, and BPAGlnl, suggests a possible role in crosslinks between desmosomal and vimentin intermediate filaments and actin. Thus, the CH domain could integrate the functions of the actin and IF cytoskeletons by physically linking these structural systems. We present an atomic model for the complex of the CH domain and actin.In crosslinking proteins that organize actin into two and three dimensional networks or gels the ABDs lie at the ends of long, flexible molecules.

scholarly journals 14-3-3 targets keratin intermediate filaments to mechanically sensitive cell–cell contacts

2020 ◽  
Vol 31 (9) ◽  
pp. 930-943 ◽  
Author(s):  
Richard A. Mariani ◽  
Shalaka Paranjpe ◽  
Radek Dobrowolski ◽  
Gregory F. Weber
Keyword(s):  
Intermediate Filaments ◽  
Intermediate Filament ◽  
Sensitive Cell ◽  
Filament Dynamics ◽  
Keratin 19 ◽  
Keratin Filaments ◽  
Novel Association ◽  
Keratin Intermediate Filaments ◽  
Cell Cell

14-3-3 serves as a major regulator of keratin intermediate filament dynamics in vivo. Migratory mesendoderm tissue of the Xenopus embryo is used to show that the dynamic reorganization of keratin filaments, a consequence of force on cell-cell adhesions, is mediated by a novel association between 14-3-3 and Keratin 19.

scholarly journals Determination of the alpha-actinin-binding site on actin filaments by cryoelectron microscopy and image analysis.

1994 ◽  
Vol 126 (2) ◽  
pp. 433-443 ◽  
Author(s):  
A McGough ◽  
M Way ◽  
D DeRosier
Keyword(s):  
Image Analysis ◽  
Smooth Muscle ◽  
Binding Sites ◽  
Actin Filaments ◽  
Three Dimensional ◽  
Actin Binding ◽  
Dimensional Structure ◽  
Outer Face ◽  
Actin Binding Domain

The three-dimensional structure of actin filaments decorated with the actin-binding domain of chick smooth muscle alpha-actinin (alpha A1-2) has been determined to 21-A resolution. The shape and location of alpha A1-2 was determined by subtracting maps of F-actin from the reconstruction of decorated filaments. alpha A1-2 resembles a bell that measures approximately 38 A at its base and extends 42 A from its base to its tip. In decorated filaments, the base of alpha A1-2 is centered about the outer face of subdomain 2 of actin and contacts subdomain 1 of two neighboring monomers along the long-pitch (two-start) helical strands. Using the atomic model of F-actin (Lorenz, M., D. Popp, and K. C. Holmes. 1993. J. Mol. Biol. 234:826-836.), we have been able to test directly the likelihood that specific actin residues, which have been previously identified by others, interact with alpha A1-2. Our results indicate that residues 86-117 and 350-375 comprise distinct binding sites for alpha-actinin on adjacent actin monomers.

scholarly journals αE-catenin actin-binding domain alters actin filament conformation and regulates binding of nucleation and disassembly factors

2013 ◽  
Vol 24 (23) ◽  
pp. 3710-3720 ◽  
Author(s):  
Scott D. Hansen ◽  
Adam V. Kwiatkowski ◽  
Chung-Yueh Ouyang ◽  
HongJun Liu ◽  
Sabine Pokutta ◽  
...  
Keyword(s):  
Actin Filament ◽  
Actin Filaments ◽  
Binding Domain ◽  
Actin Binding ◽  
Filamentous Actin ◽  
Filament Structure ◽  
Actin Binding Domain ◽  
Filament Bundles ◽  
Underlying Mechanisms ◽  
Cell Cell

The actin-binding protein αE-catenin may contribute to transitions between cell migration and cell–cell adhesion that depend on remodeling the actin cytoskeleton, but the underlying mechanisms are unknown. We show that the αE-catenin actin-binding domain (ABD) binds cooperatively to individual actin filaments and that binding is accompanied by a conformational change in the actin protomer that affects filament structure. αE-catenin ABD binding limits barbed-end growth, especially in actin filament bundles. αE-catenin ABD inhibits actin filament branching by the Arp2/3 complex and severing by cofilin, both of which contact regions of the actin protomer that are structurally altered by αE-catenin ABD binding. In epithelial cells, there is little correlation between the distribution of αE-catenin and the Arp2/3 complex at developing cell–cell contacts. Our results indicate that αE-catenin binding to filamentous actin favors assembly of unbranched filament bundles that are protected from severing over more dynamic, branched filament arrays.

scholarly journals Discovery of keratin function and role in genetic diseases: the year that 1991 was

2016 ◽  
Vol 27 (18) ◽  
pp. 2807-2810 ◽  
Author(s):  
Pierre A. Coulombe
Keyword(s):  
Intermediate Filaments ◽  
Cell Biology ◽  
Essential Role ◽  
Genetic Diseases ◽  
Structure And Function ◽  
Mechanical Support ◽  
25Th Anniversary ◽  
Keratin Filaments ◽  
And Function ◽  
Keratin Intermediate Filaments

In 1991, a set of transgenic mouse studies took the fields of cell biology and dermatology by storm in providing the first credible evidence that keratin intermediate filaments play a unique and essential role in the structural and mechanical support in keratinocytes of the epidermis. Moreover, these studies intimated that mutations altering the primary structure and function of keratin filaments underlie genetic diseases typified by cellular fragility. This Retrospective on how these studies came to be is offered as a means to highlight the 25th anniversary of these discoveries.

scholarly journals Focal segmental glomerulosclerosis ACTN4 mutants binding to actin: regulation by phosphomimetic mutations

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Hanshuang Shao ◽  
Bentley Wingert ◽  
Astrid Weins ◽  
Martin R. Pollak ◽  
Carlos Camacho ◽  
...  
Keyword(s):  
Actin Cytoskeleton ◽  
Focal Segmental Glomerulosclerosis ◽  
Actin Filaments ◽  
Binding Activity ◽  
Binding Domain ◽  
Actin Binding ◽  
Dominant Form ◽  
Cell Functions ◽  
Segmental Glomerulosclerosis ◽  
Actin Binding Domain

Abstract Natural mutations such as lysine 255 to glutamic acid (K to E), threonine 259 to isoleucine (T to I) and serine 262 to proline (S to P) that occur within the actin binding domain of alpha-actinin-4 (ACTN4) cause an autosomal dominant form of focal segmental glomerulosclerosis (FSGS) in affected humans. This appears due to elevated actin binding propensity in podocytes resulting in a ‘frozen’ cytoskeleton. What is challenging is how this cellular behavior would be compatible with other cell functions that rely on cytoskeleton plasticity. Our previous finding revealed that wild type ACTN4 can be phosphorylated at tyrosine 4 and 31 upon stimulation by epidermal growth factor (EGF) to reduce the binding to actin cytoskeleton. We queried whether the elevated actin binding activity of FSGS mutants can be downregulated by EGF-mediated phosphorylation, to discern a mechanism by which the actin-cytoskeleton can be released in FSGS. In this manuscript, we first constructed variants with Y4/31E to mimic the phosphorylation at tyrosines 4 and 31 based on earlier modeling simulations that predicted that this would bury the actin binding domains and lead to a decrease in actin binding activity. We found that Y4/31E significantly reduced the actin binding activity of K255E, T259I and S262P, dramatically preventing them from aggregating in, and inhibiting motility of, podocytes, fibroblasts and melanoma cells. A putative kinase target site at Y265 in the actin binding domain was also generated as a phosphomimetic ACTN4 Y265E that demonstrated even greater binding to actin filaments than K255E and the other FSGS mutants. That the tyrosine kinase regulation of FSGS mutation binding to actin filaments can occur in cells was shown by phosphorylation on Y4 and Y31 of the K225E after extended exposure of cells to EGF, with a decrease in ACTN4 aggregates in fibroblasts. These findings will provide evidence for targeting the N-termini of FSGS ACTN4 mutants to downregulate their actin binding activities for ameliorating the glomerulosclerotic phenotype of patients.

scholarly journals High resolution scanning electron microscopy of isolated and in situ cytoskeletal elements.

1979 ◽  
Vol 83 (1) ◽  
pp. 249-254 ◽  
Author(s):  
W Ip ◽  
D A Fischman
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
High Resolution ◽  
Intermediate Filaments ◽  
Actin Filaments ◽  
Critical Point Drying ◽  
Osmium Impregnation ◽  
Cytoskeletal Elements ◽  
Scanning Electron

Evidence is presented that cytoskeletal structures (actin filaments, intermediate filaments, and microtubules) can be resolved by scanning electron microscopy after osmium impregnation of biological material, using thiocarbohydrazide as a ligand, followed by critical-point drying. These different classes of filaments or tubules can be identified both as purified protein polymers and as structured organelles within cryofractured or detergent-extracted cells.

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