• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information

NCBI Bookshelf. A service of the National Library of Medicine, National Institutes of Health.

Cooper GM. The Cell: A Molecular Approach. 2nd edition. Sunderland (MA): Sinauer Associates; 2000.

  • By agreement with the publisher, this book is accessible by the search feature, but cannot be browsed.
Cover of The Cell

The Cell: A Molecular Approach. 2nd edition.

Show details

Intermediate Filaments

Intermediate filaments have a diameter of about 10 nm, which is intermediate between the diameters of the two other principal elements of the cytoskeleton, actin filaments (about 7 nm) and microtubules (about 25 nm). In contrast to actin filaments and microtubules, the intermediate filaments are not directly involved in cell movements. Instead, they appear to play basically a structural role by providing mechanical strength to cells and tissues.

Intermediate Filament Proteins

Whereas actin filaments and microtubules are polymers of single types of proteins (actin and tubulin, respectively), intermediate filaments are composed of a variety of proteins that are expressed in different types of cells. More than 50 different intermediate filament proteins have been identified and classified into six groups based on similarities between their amino acid sequences (Table 11.1). Types I and II consist of two groups of keratins, each consisting of about 15 different proteins, which are expressed in epithelial cells. Each type of epithelial cell synthesizes at least one type I (acidic) and one type II (neutral/basic) keratin, which copolymerize to form filaments. Some type I and II keratins (called hard keratins) are used for production of structures such as hair, nails, and horns. The other type I and II keratins (soft keratins) are abundant in the cytoplasm of epithelial cells, with different keratins being expressed in various differentiated cell types.

Table 11.1. Intermediate Filament Proteins.

Table 11.1

Intermediate Filament Proteins.

The type III intermediate filament proteins include vimentin, which is found in a variety of different kinds of cells, including fibroblasts, smooth muscle cells, and white blood cells. Another type III protein, desmin, is specifically expressed in muscle cells, where it connects the Z discs of individual contractile elements. A third type III intermediate filament protein is specifically expressed in glial cells, and a fourth is expressed in neurons of the peripheral nervous system.

The type IV intermediate filament proteins include the three neurofilament (NF) proteins (designated NF-L, NF-M, and NF-H for light, medium, and heavy, respectively). These proteins form the major intermediate filaments of many types of mature neurons. They are particularly abundant in the axons of motor neurons and are thought to play a critical role in supporting these long, thin processes, which can extend more than a meter in length. Another type IV protein (α-internexin) is expressed at an earlier stage of neuron development, prior to expression of the neurofilament proteins. The single type VI intermediate filament protein (nestin) is expressed even earlier during the development of neurons, in stem cells of the central nervous system.

The type V intermediate filament proteins are the nuclear lamins, which are found in most eukaryotic cells. Rather than being part of the cytoskeleton, the nuclear lamins are components of the nuclear envelope (see Figure 8.3). They also differ from the other intermediate filament proteins in that they assemble to form an orthogonal meshwork underlying the nuclear membrane.

Despite considerable diversity in size and amino acid sequence, the various intermediate filament proteins share a common structural organization (Figure 11.31). All of the intermediate filament proteins have a central α-helical rod domain of approximately 310 amino acids (350 amino acids in the nuclear lamins). This central rod domain is flanked by amino- and carboxy-terminal domains, which vary among the different intermediate filament proteins in size, sequence, and secondary structure. As discussed next, the α-helical rod domain plays a central role in filament assembly, while the variable head and tail domains presumably determine the specific functions of the different intermediate filament proteins.

Figure 11.31. Structure of intermediate filament proteins.

Figure 11.31

Structure of intermediate filament proteins. Intermediate filament proteins contain a central α-helical rod domain of approximately 310 amino acids (350 amino acids in the nuclear lamins). The N-terminal head and C-terminal tail domains vary in (more...)

Assembly of Intermediate Filaments

The first stage of filament assembly is the formation of dimers in which the central rod domains of two polypeptide chains are wound around each other in a coiled-coil structure, similar to that formed by myosin II heavy chains (Figure 11.32). The dimers then associate in a staggered antiparallel fashion to form tetramers, which can assemble end to end to form protofilaments. The final intermediate filament contains approximately eight protofilaments wound around each other in a ropelike structure. Because they are assembled from antiparallel tetramers, both ends of intermediate filaments are equivalent. Consequently, in contrast to actin filaments and microtubules, intermediate filaments are apolar; they do not have distinct plus and minus ends.

Figure 11.32. Assembly of intermediate filaments.

Figure 11.32

Assembly of intermediate filaments. The central rod domains of two polypeptides wind around each other in a coiled-coil structure to form dimers. Dimers then associate in a staggered antiparallel fashion to form tetramers. Tetramers associate end to end (more...)

Filament assembly requires interactions between specific types of intermediate filament proteins. For example, keratin filaments are always assembled from heterodimers containing one type I and one type II polypeptide. In contrast, the type III proteins can assemble into filaments containing only a single polypeptide (e.g., vimentin) or consisting of two different type III proteins (e.g., vimentin plus desmin). The type III proteins do not, however, form copolymers with the keratins. Among the type IV proteins, α-internexin can assemble into filaments by itself, whereas the three neurofilament proteins copolymerize to form heteropolymers.

Intermediate filaments are generally more stable than actin filaments or microtubules and do not exhibit the dynamic behavior associated with these other elements of the cytoskeleton (e.g., the treadmilling of actin filaments illustrated in Figure 11.4). However, intermediate filament proteins are frequently modified by phosphorylation, which can regulate their assembly and disassembly within the cell. The clearest example is phosphorylation of the nuclear lamins (see Figure 8.31), which results in disassembly of the nuclear lamina and breakdown of the nuclear envelope during mitosis. Cytoplasmic intermediate filaments, such as vimentin, are also phosphorylated at mitosis, which can lead to their disassembly and reorganization in dividing cells.

Intracellular Organization of Intermediate Filaments

Intermediate filaments form an elaborate network in the cytoplasm of most cells, extending from a ring surrounding the nucleus to the plasma membrane (Figure 11.33). Both keratin and vimentin filaments attach to the nuclear envelope, apparently serving to position and anchor the nucleus within the cell. In addition, intermediate filaments can associate not only with the plasma membrane but also with the other elements of the cytoskeleton, actin filaments and microtubules. Intermediate filaments thus provide a scaffold that integrates the components of the cytoskeleton and organizes the internal structure of the cell.

Figure 11.33. Intracellular organization of keratin filaments.

Figure 11.33

Intracellular organization of keratin filaments. Micrograph of epithelial cells stained with fluorescent antibodies to keratin (green). Nuclei are stained blue. The keratin filaments extend from a ring surrounding the nucleus to the plasma membrane. (Nancy (more...)

The keratin filaments of epithelial cells are tightly anchored to the plasma membrane at two areas of specialized cell contacts, desmosomes and hemidesmosomes (Figure 11.34). Desmosomes are junctions between adjacent cells, at which cell-cell contacts are mediated by transmembrane proteins related to the cadherins. On their cytoplasmic side, desmosomes are associated with a characteristic dense plaque of intracellular proteins, to which keratin filaments are attached. These attachments are mediated by desmoplakin, a member of a family of proteins called plakins that bind intermediate filaments and link them to other cellular structures. Hemidesmosomes are morphologically similar junctions between epithelial cells and underlying connective tissue, at which keratin filaments are linked by different members of the plakin family (e.g., plectin) to integrins. Desmosomes and hemidesmosomes thus anchor intermediate filaments to regions of cell-cell and cell-substratum contact, respectively, similar to the attachment of the actin cytoskeleton to the plasma membrane at adherens junctions and focal adhesions. It is important to note that the keratin filaments anchored to both sides of desmosomes serve as a mechanical link between adjacent cells in an epithelial layer, thereby providing mechanical stability to the entire tissue.

Figure 11.34. Attachment of intermediate filaments to desmosomes and hemidesmosomes.

Figure 11.34

Attachment of intermediate filaments to desmosomes and hemidesmosomes. (A) Electron micrograph illustrating keratin filaments (arrows) attached to the dense plaques of intracellular protein on both sides of a desmosome. (B) Schematic of a desmosome. Intermediate (more...)

In addition to linking intermediate filaments to cell junctions, some plakins link intermediate filaments to other elements of the cytoskeleton. Plectin, for example, binds actin filaments and microtubules in addition to intermediate filaments, so it can provide bridges between these cytoskeletal components (Figure 11.35). These bridges to intermediate filaments are thought to brace and stabilize actin filaments and microtubules, thereby increasing the mechanical stability of the cell.

Figure 11.35. Electron micrograph of plectin bridges between intermediate filaments and microtubules.

Figure 11.35

Electron micrograph of plectin bridges between intermediate filaments and microtubules. Micrograph of a fibroblast stained with antibody against plectin. The micrograph has been artificially colored to show plectin (green), antibodies against plectin (more...)

Two types of intermediate filaments, desmin and the neurofilaments, play specialized roles in muscle and nerve cells, respectively. Desmin connects the individual actin-myosin assemblies of muscle cells both to one another and to the plasma membrane, thereby linking the actions of individual contractile elements. Neurofilaments are the major intermediate filaments in most mature neurons. They are particularly abundant in the long axons of motor neurons, where they appear to be anchored to actin filaments and microtubules by neuronal members of the plakin family. Neurofilaments are thought to play an important role in providing mechanical support and stabilizing other elements of the cytoskeleton in these long, thin extensions of nerve cells.

Functions of Keratins and Neurofilaments: Diseases of the Skin and Nervous System

Although intermediate filaments have long been thought to provide structural support to the cell, direct evidence for their function has only recently been obtained. Some cells in culture make no intermediate filament proteins, indicating that these proteins are not required for the growth of cells in vitro. Similarly, injection of cultured cells with antibody against vimentin disrupts intermediate filament networks without affecting cell growth or movement. Therefore, it has been thought that intermediate filaments are most needed to strengthen the cytoskeleton of cells in the tissues of multicellular organisms, where they are subjected to a variety of mechanical stresses that do not affect cells in the isolated environment of a culture dish.

Experimental evidence for such an in vivo role of intermediate filaments was first provided in 1991 by studies in the laboratory of Elaine Fuchs. These investigators used transgenic mice to investigate the in vivo effects of expressing a keratin deletion mutant encoding a truncated polypeptide that disrupted the formation of normal keratin filaments (Figure 11.36). This mutant keratin gene was introduced into transgenic mice, where it was expressed in basal cells of the epidermis and disrupted formation of a normal keratin cytoskeleton. This resulted in the development of severe skin abnormalities, including blisters due to epidermal cell lysis following mild mechanical trauma, such as rubbing of the skin. The skin abnormalities of these transgenic mice thus provided direct support for the presumed role of keratins in providing mechanical strength to epithelial cells in tissues.

Figure 11.36. Experimental demonstration of keratin function.

Figure 11.36

Experimental demonstration of keratin function. A plasmid encoding a mutant keratin that interferes with the normal assembly of keratin filaments was microinjected into one pronucleus of a fertilized egg. Microinjected embryos were then transferred to (more...)

These experiments also pointed to the molecular basis of a human genetic disease, epidermolysis bullosa simplex (EBS). Like the transgenic mice expressing mutant keratin genes, patients with this disease develop skin blisters resulting from cell lysis after minor trauma. This similarity prompted studies of the keratin genes in EBS patients, leading to the demonstration that EBS is caused by keratin gene mutations that interfere with the normal assembly of keratin filaments. Thus, both experimental studies in transgenic mice and molecular analysis of a human genetic disease have demonstrated the role of keratins in allowing skin cells to withstand mechanical stress. Continuing studies have shown that mutations in other keratins are responsible for several other inherited skin diseases, which are similarly characterized by abnormal fragility of epidermal cells.

Other studies in transgenic mice have implicated abnormalities of neurofilaments in diseases of motor neurons, particularly amyotrophic lateral sclerosis (ALS). ALS, known as Lou Gehrig's disease and the disease afflicting the renowned physicist Stephen Hawking, results from progressive loss of motor neurons, which in turn leads to muscle atrophy, paralysis, and eventual death. ALS and other types of motor neuron disease are characterized by the accumulation and abnormal assembly of neurofilaments, suggesting that neurofilament abnormalities might contribute to these pathologies. Consistent with this possibility, overexpression of NF-L or NF-H in transgenic mice has been found to result in the development of a condition similar to ALS. Although the mechanism involved remains to be understood, these experiments clearly suggest the involvement of neurofilaments in the pathogenesis of motor neuron disease.

By agreement with the publisher, this book is accessible by the search feature, but cannot be browsed.

Copyright © 2000, Geoffrey M Cooper.
Bookshelf ID: NBK9834

Views

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...