NSP4_ROTS1
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 03-AUG-2022, entry version 120.
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; Reoviridae; Sedoreovirinae; 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.
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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:0016021; C:integral component of membrane; IEA:UniProtKB-UniRule.
DR GO; GO:0044385; C:integral to membrane of host cell; IEA:UniProtKB-KW.
DR GO; GO:0005216; F:ion channel activity; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-UniRule.
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:0039707; P:pore formation by virus in membrane of host cell; IEA:UniProtKB-KW.
DR GO; GO:0051259; P:protein complex oligomerization; IEA:UniProtKB-KW.
DR HAMAP; MF_04091; ROTA_NSP4; 1.
DR InterPro; IPR002107; Rotavirus_NSP4.
DR Pfam; PF01452; Rota_NSP4; 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
