Downloaded from http://cshperspectives.cshlp.org/ on October 4, 2021 - Published by Cold Spring Harbor Laboratory Press Cytoskeletal Integrators: The Spectrin Superfamily Ronald K.H. Liem Department of Pathology and Cell Biology, Columbia University Medical Center, New York, New York 10032 Correspondence: [email protected] SUMMARY This review discusses the spectrin superfamily of proteins that function to connect cytoskeletal elements to each other, the cell membrane, and the nucleus. The signature domain is the spectrin repeat, a 106–122-amino-acid segment comprising three a-helices. a-actinin is considered to be the ancestral protein and functions to cross-link actin filaments. It then evolved to generate spectrin and dystrophin that function to link the actin cytoskeleton to the cell membrane, as well as the spectraplakins and plakins that link cytoskeletal elements to each other and to junctional complexes. A final class comprises the nesprins, which are able to bind to the nuclear membrane. This review discusses the domain organization of the various spectrin family members, their roles in protein–protein interactions, and their roles in disease, as determined from mutations, and it also describes the functional roles of the family members as determined from null phenotypes. Outline 1 Introduction 5 Spectraplakins and plakins 2 a-actinin 6 Nesprins 3 Spectrins 7 Concluding remarks 4 Dystrophin and utrophin References Editors: Thomas D. Pollard and Robert D. Goldman Additional Perspectives on The Cytoskeleton available at www.cshperspectives.org Copyright # 2016 Cold Spring Harbor Laboratory Press; all rights reserved; doi: 10.1101/cshperspect.a018259 Cite this article as Cold Spring Harb Perspect Biol 2016;8:a018259 1 Downloaded from http://cshperspectives.cshlp.org/ on October 4, 2021 - Published by Cold Spring Harbor Laboratory Press R.K.H. Liem 1 INTRODUCTION fore considering the spectraplakin/plakin members, and finally the nesprin proteins. Cytoskeletal elements, such as microtubules, microfila- ments, and intermediate filaments, are linked to each other, 2 a-ACTININ to the cell membrane, and to the nucleus by a class of cytoskeletal integrators. The common feature among these a-actinin is known primarily as an actin-cross-linking pro- cytoskeletal integrators is the presence of tandem arrays of tein and has been reviewed in this capacity elsewhere (Pol- units known as spectrin repeats (SRs). In addition, many of lard 2016). It is included in this review of cytoskeletal these proteins have calponin-homology (CH) domains integrators because it appears to be the ancestor protein that bind to actin filaments. Other features found in vari- to the family of proteins described here (Broderick and ous subclasses of these cytoskeletal integrators are domains Winder 2005). a-actinin comprises three domains, an actin that bind to microtubules, intermediate filaments, and the filament–binding domain consisting of two CH domains, nuclear membrane. Furthermore, there are other protein– a series of four SRs, and a domain consisting of EF hand– protein interaction domains in most of these molecules. like motifs (see Fig. 1A). As these domains are also present First, I will discuss the simplest of these proteins—a-acti- in the other cytoskeletal integrators discussed herein, they nin—followed by spectrin and dystrophin/utrophin, be- are described in more detail below. α-actinin A KEY CH1 CH2 EF CH1CH2 Actin-binding domain PH domain EF CH2 CH1 EF EF-hand motif CC-ROD β -spectrin Spectrin repeats Plakin repeat CH1 CH2 W WW domain GAR domain EF Coiled coils GSR domain α-spectrin ZZ ZZ domain Plakin-like repeat Dystrophin SH3 domain KASH domain CH1 CH2 W EF ZZ Utrophin CH1 CH2 W EF ZZ B Shot I CH1 CH2 EF Shot II CH1 CH2 EF VAB-10A CH1 CH2 VAB-10B CH1 CH2 EF Figure 1. Schematic diagrams of the spectrin superfamily of cytoskeletal linker proteins. (A) a-actinin, spectrin, and dystrophin/utrophin. a-actinin is shown as an antiparallel dimer. The two actin-binding domains (ABDs) CH1 and CH2 allow the cross-linking of actin filaments. Spectrin is shown as a dimer of a- and b-spectrin, with a-spectrin shown below b-spectrin in an antiparallel orientation (from carboxyl to amino terminus). A tetramer can be formed by a second antiparallel dimer in the opposite direction, with the amino terminus of a-spectrin interacting with the carboxyl terminus of b-spectrin (Broderick and Winder 2005). The resulting tetramer resembles a-actinin, but has many more spectrin repeats (SRs). Dystrophin is shown as a monomer. It can interact with actin through the ABD, but has many more interaction domains, including a second ABD in the last two SRs. Utrophin is similar to dystrophin, except that the last two SRs are missing. (Figure and legend continue on facing page.) 2 Cite this article as Cold Spring Harb Perspect Biol 2016;8:a018259 Downloaded from http://cshperspectives.cshlp.org/ on October 4, 2021 - Published by Cold Spring Harbor Laboratory Press The Spectrin Superfamily 2.1 CH Domains nin, where it was identified before it was found to be in actin-binding proteins. The CH1 domain is able to bind to The actin-binding domain (ABD) of the spectrin family of actin filaments by itself, albeit with lower affinity than the proteins consists of two calponin-homology (CH1 and combined CH1–CH2 domains (Gimona and Winder CH2) domains in tandem. These ABDs are found near 1998). In contrast, the CH2 domain alone cannot bind to the amino termini of these proteins. Calponin, which is a actin filaments. None of the members of the spectrin super- calcium-binding protein present in smooth muscle, has a family expresses only the CH1 domain, although there are single CH domain that is not able to bind to actin. It is the isoforms that express only the CH2 domain (Jefferson et al. prototype CH domain because it was first found in calpo- 2004). Structurally, the CH domain is a compact globular C MACF1a CH1 CH2 EF MACF1b CH1 CH2 A A C b c EF BPAG1a CH1 CH2 EF BPAG1b CH1 CH2 A EF BPAG1e B C Plectin CH1 CH2 B B B B B C Desmoplakin I A B C D Nesprin-1G CH2CH1 Nesprin-2G CH1 CH2 Nesprin-3α Nesprin-4 Figure 1. (Continued) The domain labeled “ZZ” is the zinc-finger domain. (B) Invertebrate spectraplakins. The two splice forms of the two known invertebrate spectraplakins are shown. Shots I and II are the Drosophila spectraplakins, and VAB-10aand VAB-10bare the Caenorhabditis elegans spectraplakins. The identityof the various domains is given in the key. (C) Vertebrate spectraplakins and plakins. The major “a” and “b” splice forms of microtubule-actin cross- linking factor 1 (MACF1) and bullous pemphigoid antigen 1 (BPAG1) are shown. MACF1a and MACF1b have domain structures similar to those of BPAG1a and BPAG1b, respectively. However, a third splice form of BPAG1— BPAG1e—only shares the so-called plakin domain in common with the other two isoforms. BPAG1e can dimerize through its coiled-coil rod. Plectin and desmoplakin, along with BPAG1e, are considered “plakins” rather than “spectraplakins,” although the plakin domain actually consists of SRs. (D) Nesprins. The four major isoforms of the nesprins are shown. There are some unusual SRs in the two giant isoforms, but, for simplicity, they are shown here as standard SRs. Cite this article as Cold Spring Harb Perspect Biol 2016;8:a018259 3 Downloaded from http://cshperspectives.cshlp.org/ on October 4, 2021 - Published by Cold Spring Harbor Laboratory Press R.K.H. Liem domain that comprises four a-helices that are 11–18 res- idues in length and show a roughly parallel orientation, interspersed with three shorter helices (Broderick and A Winder 2002). 2.2 SR Domains C Repeat 1 SRs occur in tandem and constitute the so-called rod do- main or SR domain. This domain is required for proteins to N be considered members of the spectrin superfamily of cy- L toskeletal integrators. These SR domains consist of between B 2 and 74 SRs and act as flexible spacers between the actin E cytoskeleton and the membrane or other cytoskeletal ele- Repeat 2 ments. In addition, the SR domain can function to interact D with other molecules in the cytoplasm. The repeats are 106–122 residues in length. Each repeat comprises three a-helices, and tandem SRs are connected with an inter- spectrin repeat that is also helical in nature (Fig. 2). F Proteins with spectrin repeat–containing rod domains C can dimerize in either a parallel or antiparallel manner. SRs Figure 2. Ribbon diagram illustrating the structure of two spectrin can have limited homology at the amino acid level but are repeats (SRs) in the plakin domain of bullous pemphigoid antigen 1 related structurally. Protein-prediction programs can iden- (BPAG1/dystonin), which is a member of the spectrin superfamily. tify SRs based on the expected secondary structure. Depicted is a loop-like region (green) at the far amino-terminal end, followed by a pair of SRs 1 and 2 (red and blue, respectively) arranged in tandem and connected by a linker region (L, purple) that is also 2.3 EF Hands helical in nature. Also labeled are the component helices A–C and D–F. (Reprinted from Jefferson et al. 2004.) The carboxyl terminus of a-actinin has an EF hand motif. The EF hand consists of two a-helices linked bya short loop found to be a common component that lines the intra- region. This helix–loop–helix arrangement resembles the cellular side of the plasma membrane of cells of many ver- spread thumb and forefinger of a hand. Calcium ions can tebrate tissues and is also present in a number of bind to the EF hand, leading to a conformational change nonvertebrates.