ID NSP4_ROTS1 Reviewed; 175 AA. AC P04512; DT 13-AUG-1987, integrated into UniProtKB/Swiss-Prot. DT 13-AUG-1987, sequence version 1. DT 24-JAN-2024, entry version 124. DE RecName: Full=Non-structural glycoprotein 4 {ECO:0000255|HAMAP-Rule:MF_04091}; DE Short=NSP4 {ECO:0000255|HAMAP-Rule:MF_04091}; DE AltName: Full=NCVP5 {ECO:0000255|HAMAP-Rule:MF_04091}; DE AltName: Full=NS28 {ECO:0000255|HAMAP-Rule:MF_04091}; OS Rotavirus A (strain RVA/SA11-Both/G3P5B[2]) (RV-A) (Simian Agent 11 (strain OS Both)). OC Viruses; Riboviria; Orthornavirae; Duplornaviricota; Resentoviricetes; OC Reovirales; Sedoreoviridae; Rotavirus; Rotavirus A. OX NCBI_TaxID=37137; OH NCBI_TaxID=9544; Macaca mulatta (Rhesus macaque). RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC RNA], SUBCELLULAR LOCATION, STRUCTURE OF RP CARBOHYDRATE, AND GLYCOSYLATION AT ASN-8 AND ASN-18. RX PubMed=6312090; DOI=10.1128/jvi.48.2.335-339.1983; RA Both G.W., Siegman L.J., Bellamy A.R., Atkinson P.H.; RT "Coding assignment and nucleotide sequence of simian rotavirus SA11 gene RT segment 10: location of glycosylation sites suggests that the signal RT peptide is not cleaved."; RL J. Virol. 48:335-339(1983). RN [2] RP PROTEIN SEQUENCE OF 112-120, PROTEOLYTIC CLEAVAGE, AND FUNCTION. RX PubMed=11090165; DOI=10.1128/jvi.74.24.11663-11670.2000; RA Zhang M., Zeng C.Q.-Y., Morris A.P., Estes M.K.; RT "A functional NSP4 enterotoxin peptide secreted from rotavirus-infected RT cells."; RL J. Virol. 74:11663-11670(2000). RN [3] RP FUNCTION, SUBUNIT, SUBCELLULAR LOCATION, AND MUTAGENESIS OF CYS-63; CYS-71; RP MET-175 AND 174-ALA--MET-175. RX PubMed=1316468; DOI=10.1128/jvi.66.6.3566-3572.1992; RA Taylor J.A., Meyer J.C., Legge M.A., O'Brien J.A., Street J.E., Lord V.J., RA Bergmann C.C., Bellamy A.R.; RT "Transient expression and mutational analysis of the rotavirus RT intracellular receptor: the C-terminal methionine residue is essential for RT ligand binding."; RL J. Virol. 66:3566-3572(1992). RN [4] RP FUNCTION. RX PubMed=7637021; DOI=10.1128/jvi.69.9.5763-5772.1995; RA Tian P., Estes M.K., Hu Y., Ball J.M., Zeng C.Q.-Y., Schilling W.P.; RT "The rotavirus nonstructural glycoprotein NSP4 mobilizes Ca2+ from the RT endoplasmic reticulum."; RL J. Virol. 69:5763-5772(1995). RN [5] RP COILED-COIL DOMAIN. RX PubMed=8887538; DOI=10.1002/j.1460-2075.1996.tb00824.x; RA Taylor J.A., O'Brien J.A., Yeager M.; RT "The cytoplasmic tail of NSP4, the endoplasmic reticulum-localized non- RT structural glycoprotein of rotavirus, contains distinct virus binding and RT coiled coil domains."; RL EMBO J. 15:4469-4476(1996). RN [6] RP FUNCTION. RX PubMed=9108087; DOI=10.1073/pnas.94.8.3960; RA Dong Y., Zeng C.Q.-Y., Ball J.M., Estes M.K., Morris A.P.; RT "The rotavirus enterotoxin NSP4 mobilizes intracellular calcium in human RT intestinal cells by stimulating phospholipase C-mediated inositol 1,4,5- RT trisphosphate production."; RL Proc. Natl. Acad. Sci. U.S.A. 94:3960-3965(1997). RN [7] RP INTERACTION WITH MICROTUBULES, AND SUBCELLULAR LOCATION. RX PubMed=11101519; DOI=10.1093/emboj/19.23.6465; RA Xu A., Bellamy A.R., Taylor J.A.; RT "Immobilization of the early secretory pathway by a virus glycoprotein that RT binds to microtubules."; RL EMBO J. 19:6465-6474(2000). RN [8] RP INTERACTION WITH ICP, AND MUTAGENESIS OF TYR-166. RX PubMed=10799621; DOI=10.1128/jvi.74.11.5388-5394.2000; RA O'Brien J.A., Taylor J.A., Bellamy A.R.; RT "Probing the structure of rotavirus NSP4: a short sequence at the extreme C RT terminus mediates binding to the inner capsid particle."; RL J. Virol. 74:5388-5394(2000). RN [9] RP ER RETENTION SIGNAL. RX PubMed=12655088; DOI=10.1099/vir.0.18786-0; RA Mirazimi A., Magnusson K.-E., Svensson L.; RT "A cytoplasmic region of the NSP4 enterotoxin of rotavirus is involved in RT retention in the endoplasmic reticulum."; RL J. Gen. Virol. 84:875-883(2003). RN [10] RP INTERACTION WITH HUMAN CAV1, AND SUBCELLULAR LOCATION. RX PubMed=16501093; DOI=10.1128/jvi.80.6.2842-2854.2006; RA Parr R.D., Storey S.M., Mitchell D.M., McIntosh A.L., Zhou M., Mir K.D., RA Ball J.M.; RT "The rotavirus enterotoxin NSP4 directly interacts with the caveolar RT structural protein caveolin-1."; RL J. Virol. 80:2842-2854(2006). RN [11] RP SUBCELLULAR LOCATION. RX PubMed=16731945; DOI=10.1128/jvi.02167-05; RA Berkova Z., Crawford S.E., Trugnan G., Yoshimori T., Morris A.P., RA Estes M.K.; RT "Rotavirus NSP4 induces a novel vesicular compartment regulated by calcium RT and associated with viroplasms."; RL J. Virol. 80:6061-6071(2006). RN [12] RP SUBCELLULAR LOCATION, AND FUNCTION. RX PubMed=17035333; DOI=10.1128/jvi.01378-06; RA Bugarcic A., Taylor J.A.; RT "Rotavirus nonstructural glycoprotein NSP4 is secreted from the apical RT surfaces of polarized epithelial cells."; RL J. Virol. 80:12343-12349(2006). RN [13] RP INTERACTION WITH HUMAN CAV1, SUBCELLULAR LOCATION, AND GLYCOSYLATION. RC STRAIN=SA11-4F; RX PubMed=17376898; DOI=10.1128/jvi.01862-06; RA Storey S.M., Gibbons T.F., Williams C.V., Parr R.D., Schroeder F., RA Ball J.M.; RT "Full-length, glycosylated NSP4 is localized to plasma membrane caveolae by RT a novel raft isolation technique."; RL J. Virol. 81:5472-5483(2007). RN [14] RP INTERACTION WITH HUMAN INTEGRIN ITGA1/ITGB1 HETERODIMER, INTERACTION WITH RP HUMAN INTEGRIN ITGA2/ITGB1 HETERODIMER, AND MUTAGENESIS OF GLU-120. RX PubMed=18587047; DOI=10.1073/pnas.0803934105; RA Seo N.-S., Zeng C.Q.-Y., Hyser J.M., Utama B., Crawford S.E., Kim K.J., RA Hoeoek M., Estes M.K.; RT "Inaugural article: integrins alpha1beta1 and alpha2beta1 are receptors for RT the rotavirus enterotoxin."; RL Proc. Natl. Acad. Sci. U.S.A. 105:8811-8818(2008). RN [15] RP FUNCTION, AND SUBCELLULAR LOCATION. RX PubMed=21151776; DOI=10.1128/mbio.00265-10; RA Hyser J.M., Collinson-Pautz M.R., Utama B., Estes M.K.; RT "Rotavirus disrupts calcium homeostasis by NSP4 viroporin activity."; RL MBio 1:0-0(2010). RN [16] RP X-RAY CRYSTALLOGRAPHY (1.86 ANGSTROMS) OF 95-137 IN COMPLEX WITH CALCIUM OR RP STRONTIUM, METAL-BINDING AT GLN-123 AND GLU-120, AND SUBUNIT. RX PubMed=11124032; DOI=10.1006/jmbi.2000.4250; RA Bowman G.D., Nodelman I.M., Levy O., Lin S.L., Tian P., Zamb T.J., RA Udem S.A., Venkataraghavan B., Schutt C.E.; RT "Crystal structure of the oligomerization domain of NSP4 from rotavirus RT reveals a core metal-binding site."; RL J. Mol. Biol. 304:861-871(2000). CC -!- FUNCTION: Plays an essential role in the virus replication cycle by CC acting as a viroporin. Creates a pore in the host reticulum endoplasmic CC and as a consequence releases Ca(2+) in the cytoplasm of infected cell. CC In turn, high levels of cytoplasmic calcium trigger membrane CC trafficking and transport of viral ER-associated proteins to CC viroplasms, sites of viral genome replication and immature particle CC assembly. {ECO:0000255|HAMAP-Rule:MF_04091, CC ECO:0000269|PubMed:21151776, ECO:0000269|PubMed:7637021, CC ECO:0000269|PubMed:9108087}. CC -!- FUNCTION: The secreted form acts as an enterotoxin that causes CC phospholipase C-dependent elevation of the intracellular calcium CC concentration in host intestinal mucosa cells. Increased concentration CC of intracellular calcium disrupts the cytoskeleton and the tight CC junctions, raising the paracellular permeability. Potentiates chloride CC ion secretion through a calcium ion-dependent signaling pathway, CC inducing age-dependent diarrhea. To perform this enterotoxigenic role CC in vivo, NSP4 is released from infected enterocytes in a soluble form CC capable of diffusing within the intestinal lumen and interacting with CC host plasma membrane receptors on neighboring epithelial cells such as CC integrins ITGA1/ITGB1 and ITGA2/ITGB1. {ECO:0000255|HAMAP- CC Rule:MF_04091, ECO:0000269|PubMed:17035333, CC ECO:0000269|PubMed:18587047}. CC -!- SUBUNIT: Homotetramer. Interacts with the immature particle in the CC viroplasm. Interacts with host CAV1, early and late in infection. CC Interacts with host integrin ITGA1/ITGB1 heterodimer. Interacts with CC host integrin ITGA2/ITGB1 heterodimer (PubMed:18587047). Interaction CC with microtubules blocks trafficking to the Golgi apparatus CC (PubMed:11101519). {ECO:0000255|HAMAP-Rule:MF_04091, CC ECO:0000269|PubMed:10799621, ECO:0000269|PubMed:11101519, CC ECO:0000269|PubMed:11124032, ECO:0000269|PubMed:1316468, CC ECO:0000269|PubMed:16501093, ECO:0000269|PubMed:17376898, CC ECO:0000269|PubMed:18587047}. CC -!- INTERACTION: CC P04512; P56199: ITGA1; Xeno; NbExp=2; IntAct=EBI-15711650, EBI-2554465; CC P04512; P17301: ITGA2; Xeno; NbExp=3; IntAct=EBI-15711650, EBI-702960; CC -!- SUBCELLULAR LOCATION: Host rough endoplasmic reticulum membrane CC {ECO:0000250|UniProtKB:P08434, ECO:0000255|HAMAP-Rule:MF_04091}; CC Single-pass type III membrane protein {ECO:0000255|HAMAP- CC Rule:MF_04091}. Host membrane, host caveola {ECO:0000255|HAMAP- CC Rule:MF_04091, ECO:0000269|PubMed:17376898}; Single-pass type III CC membrane protein {ECO:0000255|HAMAP-Rule:MF_04091}. Secreted CC {ECO:0000255|HAMAP-Rule:MF_04091, ECO:0000269|PubMed:17035333}. CC Note=NSP4 localizes also in vesicular structures which contain CC autophagosomal markers and associate with viroplasms in virus-infected CC cells. Additionally, a soluble form of glycosylated NSP4 is secreted CC despite retention of its transmembrane domain. {ECO:0000255|HAMAP- CC Rule:MF_04091}. CC -!- DOMAIN: Binds 1 calcium ion per tetramer. {ECO:0000255|HAMAP- CC Rule:MF_04091}. CC -!- DOMAIN: The coiled coil region mediates oligomerization. CC -!- PTM: The N-glycosyl content is primarily Man(9)GlcNAc, with a small CC amount of Man(8)GlcNAc. {ECO:0000255|HAMAP-Rule:MF_04091}. CC -!- SIMILARITY: Belongs to the rotavirus NSP4 family. {ECO:0000255|HAMAP- CC Rule:MF_04091}. CC -!- CAUTION: A candidate enterotoxigenic cleaved form of the protein has CC been suggested [PubMed:11090165], but it remains unclear whether this CC truncated form constitutes an active enterotoxin in vivo. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; K01138; AAA47291.1; -; Genomic_RNA. DR PIR; A04140; VGXRTS. DR PDB; 1G1I; X-ray; 2.00 A; A/B=95-137. DR PDB; 1G1J; X-ray; 1.86 A; A/B=95-137. DR PDB; 2O1K; X-ray; 1.67 A; A/B=95-146. DR PDBsum; 1G1I; -. DR PDBsum; 1G1J; -. DR PDBsum; 2O1K; -. DR SMR; P04512; -. DR DIP; DIP-46136N; -. DR IntAct; P04512; 2. DR TCDB; 1.A.94.1.1; the rotavirus non-structural glycoprotein 4 viroporin (nsp4) family. DR iPTMnet; P04512; -. DR EvolutionaryTrace; P04512; -. DR Proteomes; UP000007180; Genome. DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell. DR GO; GO:0044155; C:host caveola; IEA:UniProtKB-SubCell. DR GO; GO:0044169; C:host cell rough endoplasmic reticulum membrane; IEA:UniProtKB-SubCell. DR GO; GO:0016020; C:membrane; IEA:UniProtKB-UniRule. DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-UniRule. DR GO; GO:0005216; F:monoatomic ion channel activity; IEA:UniProtKB-KW. DR GO; GO:0090729; F:toxin activity; IEA:UniProtKB-UniRule. DR GO; GO:0039520; P:induction by virus of host autophagy; IEA:UniProtKB-KW. DR GO; GO:0051259; P:protein complex oligomerization; IEA:UniProtKB-KW. DR GO; GO:0039707; P:virus-mediated pore formation in host cell membrane; IEA:UniProtKB-KW. DR Gene3D; 1.20.5.430; -; 1. DR HAMAP; MF_04091; ROTA_NSP4; 1. DR InterPro; IPR002107; Rotavirus_NSP4. DR Pfam; PF01452; Rota_NSP4; 1. DR SUPFAM; SSF58030; Rotavirus nonstructural proteins; 1. PE 1: Evidence at protein level; KW 3D-structure; Activation of host autophagy by virus; Calcium; KW Direct protein sequencing; Enterotoxin; Glycoprotein; KW Host endoplasmic reticulum; Host membrane; Host-virus interaction; KW Ion channel; Ion transport; Membrane; Metal-binding; Reference proteome; KW Secreted; Signal-anchor; Toxin; Transmembrane; Transmembrane helix; KW Transport; Viral ion channel; Virulence. FT CHAIN 1..175 FT /note="Non-structural glycoprotein 4" FT /id="PRO_0000149628" FT TOPO_DOM 1..28 FT /note="Lumenal" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04091" FT TRANSMEM 29..51 FT /note="Helical; Signal-anchor for type III membrane FT protein" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04091" FT TOPO_DOM 52..175 FT /note="Cytoplasmic" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04091" FT REGION 122..175 FT /note="Required for interaction with microtubules" FT REGION 159..175 FT /note="ICP binding domain" FT BINDING 120 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_note="ligand shared between all tetrameric FT partners" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04091" FT BINDING 123 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_note="ligand shared between all tetrameric FT partners" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04091" FT CARBOHYD 8 FT /note="N-linked (GlcNAc...) asparagine; by host" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04091, FT ECO:0000269|PubMed:6312090" FT CARBOHYD 18 FT /note="N-linked (GlcNAc...) asparagine; by host" FT /evidence="ECO:0000255|HAMAP-Rule:MF_04091, FT ECO:0000269|PubMed:6312090" FT MUTAGEN 63 FT /note="C->S: No effect on receptor activity nor on oligomer FT formation. No effect on receptor activity nor on oligomer FT formation; when associated with C-71." FT /evidence="ECO:0000269|PubMed:1316468" FT MUTAGEN 71 FT /note="C->S: No effect on receptor activity nor on oligomer FT formation. No effect on receptor activity nor on oligomer FT formation; when associated with C-63." FT /evidence="ECO:0000269|PubMed:1316468" FT MUTAGEN 120 FT /note="E->A: Complete loss of interaction with integrin FT ITGA2." FT /evidence="ECO:0000269|PubMed:18587047" FT MUTAGEN 166 FT /note="Y->I: Loss of ICP-binding activity." FT /evidence="ECO:0000269|PubMed:10799621" FT MUTAGEN 174..175 FT /note="Missing: Abolishes receptor activity." FT /evidence="ECO:0000269|PubMed:1316468" FT MUTAGEN 175 FT /note="M->G,K,I: Abolishes receptor activity." FT /evidence="ECO:0000269|PubMed:1316468" FT MUTAGEN 175 FT /note="Missing: Abolishes receptor activity. No effect on FT subcellular location." FT /evidence="ECO:0000269|PubMed:1316468" FT HELIX 96..136 FT /evidence="ECO:0007829|PDB:2O1K" SQ SEQUENCE 175 AA; 20267 MW; 1E72A3223C567309 CRC64; MEKLTDLNYT LSVITLMNNT LHTILEDPGM AYFPYIASVL TGLFALNKAS IPTMKIALKT SKCSYKVVKY CIVTIFNTLL KLAGYKEQIT TKDEIEKQMD RVVKEMRRQL EMIDKLTTRE IEQVELLKRI YDKLTVQTTG EIDMTKEINQ KNVRTLEEWE SGKNPYEPRE VTAAM //