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3.A.27.  The Endoplasmic Reticulum Membrane Protein Insertion Complex (EMC) Family

Guna et al. 2018 found that known membrane insertion pathways fail to effectively engage tail-anchored membrane proteins with moderately hydrophobic C-terminal transmembrane domains. These proteins are instead shielded in the cytosol by calmodulin. Dynamic release from calmodulin allowed sampling of the endoplasmic reticulum (ER), where the conserved ER membrane protein complex (EMC) was shown to be essential for efficient insertion in vitro and in cells. Purified EMC in synthetic liposomes catalyzed the insertion of its substrates in a reconstituted system. Thus, EMC is a transmembrane domain insertase, a function that may explain its widely pleiotropic membrane-associated phenotypes across organisms (Guna et al. 2018). This family can be considered to be the second C-terminal Tail-Anchored Membrane Protein Biogenesis/Insertion Complex Family, the first having TC# 3.A.21. This system has been reviewed (Volkmar and Christianson 2020).

The ten known proteins of the EMC are EMC1 - EMC10.  These proteins have sizes between 110 and 993 aas, seven of them having sizes between 180 and 300 aas in humans (see 3.A.27.1.1).  These proteins have between ) and 3 TMSs each.  None of them are clearly homologous to any of the proteins in TCDB based on sequence similarity.  However, Guna et al. 2018 reported that EMC3 is distantly related to the yeast Get1 protein and the mammalian TRC40 protein, subunits of the insertase for the TRC pathway (Wang et al. 2014) (see TC#s 3.A.19 and 3.A.21). Get1 and EMC3 may have evolved from the ancestral prokaryotic insertase of the YidC family as claimed by Guna et al., 2018,  The EMC has been genetically implicated in several membrane-associated processes such as quality control, trafficking, protein maturation, and lipid homeostasis (Jonikas et al. 2009; Christianson et al. 2011). 

High-throughput genetic interaction analyses have revealed the EMC, and its disruption has since been found to affect wide-ranging processes, including protein trafficking, organelle communication, ER stress, viral maturation, lipid homeostasis, integral membrane insertion and folding, and others (Chitwood and Hegde 2019).  Biochemical reconstitution experiments showed that EMC can directly mediate the insertion of TMSs into the lipid bilayer. A central role for EMC as a TMD insertion factor explains its high abundance, wide conservation, and pleiotropic phenotypes. Tian et al. 2019 identified 36 EMC-dependent membrane proteins and 171 EMC- independent membrane proteins, and found that a common feature among EMC-dependent proteins is the occurance of TMSs with polar and/or charged residues; the EMC may be involved in handling TMDs with residues challenging for membrane integration.

Hiramatsu et al. 2019 investigated the EMC function using the Drosophila photoreceptor as a model system. EMC was necessary only for the biogenesis of a subset of multi-pass membrane proteins such as rhodopsin (Rh1), TRP, TRPL, Csat, Cni, SERCA, and Na+K+ATPase alpha, but not for that of secretory or single-pass membrane proteins. In EMC-deficient cells, Rh1 was translated to its C-terminus but degraded independently from ER-associated degradation. Thus, EMC exerts its effect after translation but during the membrane integration of TMDs. EMC is not required for the stable expression of the first three TMSs of Rh1 but is required for that of the fourth and fifth TMSs. EMC is required for the ER membrane insertion of succeeding TMDs of multi-pass membrane proteins (Hiramatsu et al. 2019). 

O'Donnell et al. 2020 defined the composition and architecture of human EMC. EMC's cytosolic domain contains a large, moderately hydrophobic vestibule that can bind a substrate's TMS. The cytosolic vestibule leads into a lumenally-sealed, lipid-exposed, intramembrane groove large enough to accommodate a single substrate TMS. A gap between the cytosolic vestibule and the intramembrane groove provides a potential path for substrate egress from EMC. Thus, EMC facilitates energy-independent membrane insertion of TMSs, explain why only short lumenal domains are translocated by EMC, and constrains models of EMC's proposed chaperone function (O'Donnell et al. 2020).

The nine- or ten-subunit endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved co- and posttranslational insertase at the ER. Pleiner et al. 2020 determined the structure of the human EMC in a lipid nanodisc to an overall resolution of 3.4 Å by cryo-EM, permitting building of a nearly complete atomic model. They used structure-guided mutagenesis to demonstrate that substrate insertion requires a methionine-rich cytosolic loop and occurs via an enclosed hydrophilic vestibule within the membrane formed by the subunits EMC3 and EMC6. The EMC may use local membrane thinning and a positively charged patch to decrease the energetic barrier for insertion into the bilayer (Pleiner et al. 2020).

The reaction belived to be catalyzed by the EMC complex is:

C-terminal tail anchored protein with a moderately hydrophobic TMS (associated with calmodulin in the cytoplasm) → C-terminal tail anchored protein with a moderately hydrophobic TMS (anchored to the ER membrane).

 

 

References associated with 3.A.27 family:

Bai, L., Q. You, X. Feng, A. Kovach, and H. Li. (2020). Structure of the ER membrane complex, a transmembrane-domain insertase. Nature. [Epub: Ahead of Print] 32494008
Chitwood, P.J. and R.S. Hegde. (2019). The Role of EMC during Membrane Protein Biogenesis. Trends Cell Biol. 29: 371-384. 30826214
Chitwood, P.J., S. Juszkiewicz, A. Guna, S. Shao, and R.S. Hegde. (2018). EMC Is Required to Initiate Accurate Membrane Protein Topogenesis. Cell 175: 1507-1519.e16. 30415835
Christianson, J.C., J.A. Olzmann, T.A. Shaler, M.E. Sowa, E.J. Bennett, C.M. Richter, R.E. Tyler, E.J. Greenblatt, J.W. Harper, and R.R. Kopito. (2011). Defining human ERAD networks through an integrative mapping strategy. Nat. Cell Biol. 14: 93-105. 22119785
Guna, A., N. Volkmar, J.C. Christianson, and R.S. Hegde. (2018). The ER membrane protein complex is a transmembrane domain insertase. Science 359: 470-473. 29242231
Hiramatsu, N., T. Tago, T. Satoh, and A.K. Satoh. (2019). ER membrane protein complex is required for the insertions of late-synthesized transmembrane helices of Rh1 in photoreceptors. Mol. Biol. Cell mbcE19080434. [Epub: Ahead of Print] 31553680
Jonikas, M.C., S.R. Collins, V. Denic, E. Oh, E.M. Quan, V. Schmid, J. Weibezahn, B. Schwappach, P. Walter, J.S. Weissman, and M. Schuldiner. (2009). Comprehensive characterization of genes required for protein folding in the endoplasmic reticulum. Science 323: 1693-1697. 19325107
Marquez, J., J. Criscione, R.M. Charney, M.S. Prasad, W.Y. Hwang, E.K. Mis, M.I. GarcĂ­a-Castro, and M.K. Khokha. (2020). Disrupted ER membrane protein complex-mediated topogenesis drives congenital neural crest defects. J Clin Invest. [Epub: Ahead of Print] 31904590
O''Donnell, J.P., B.P. Phillips, Y. Yagita, S. Juszkiewicz, A. Wagner, D. Malinverni, R.J. Keenan, E.A. Miller, and R.S. Hegde. (2020). The architecture of EMC reveals a path for membrane protein insertion. Elife 9:. 32459176
Pleiner, T., G.P. Tomaleri, K. Januszyk, A.J. Inglis, M. Hazu, and R.M. Voorhees. (2020). Structural basis for membrane insertion by the human ER membrane protein complex. Science 369: 433-436. 32439656
Tian, S., Q. Wu, B. Zhou, M.Y. Choi, B. Ding, W. Yang, and M. Dong. (2019). Proteomic Analysis Identifies Membrane Proteins Dependent on the ER Membrane Protein Complex. Cell Rep 28: 2517-2526.e5. 31484065
Volkmar, N. and J.C. Christianson. (2020). Squaring the EMC - how promoting membrane protein biogenesis impacts cellular functions and organismal homeostasis. J Cell Sci 133:. 32332093
Wang, F., C. Chan, N.R. Weir, and V. Denic. (2014). The Get1/2 transmembrane complex is an endoplasmic-reticulum membrane protein insertase. Nature 512: 441-444. 25043001