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P04512 (NSP4_ROTS1) Reviewed, UniProtKB/Swiss-Prot

Last modified February 19, 2014. Version 83. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (2) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
Non-structural glycoprotein 4

Short name=NSP4
Alternative name(s):
NCVP5
NS28
OrganismRotavirus A (strain SA11-Both G3-P5B[2]-I2-R2-C5-M5-A5-N2-T5-E2-H5) (RV-A) (Simian Agent 11 (strain Both)) [Complete proteome]
Taxonomic identifier37137 [NCBI]
Taxonomic lineageVirusesdsRNA virusesReoviridaeSedoreovirinaeRotavirusRotavirus A
Virus hostMacaca mulatta (Rhesus macaque) [TaxID: 9544]

Protein attributes

Sequence length175 AA.
Sequence statusComplete.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Involved in virus morphogenesis. Functions as a receptor for the immature double-layered inner capsid particle (ICP) which transiently buds into the lumen of the rough endoplasmic reticulum during viral maturation. Ref.2 Ref.3 Ref.4 Ref.6

Enterotoxin that causes a phospholipase C-dependent elevation of the intracellular calcium concentration in host intestinal mucosa cells. Increased concentration of intracellular calcium disrupts the cytoskeleton and the tight junctions, raising the paracellular permeability. Potentiates chloride ion secretion through a calcium ion-dependent signaling pathway, inducing age-dependent diarrhea. To perform this enterotoxigenic role in vivo, NSP4 is probably released from infected enterocytes in a soluble form capable of diffusing within the intestinal lumen and interacting with the plasma membrane receptors on neighboring epithelial cells. Possible receptors for NSP4 are alpha-1/beta-1 and alpha-2/beta-1 integrin heterodimers. Ref.2 Ref.3 Ref.4 Ref.6

Subunit structure

Homotetramer. Forms a complex with the ICP. Interacts, via the active enterotoxic peptide region, with host CAV1, early and late in infection. Interacts with host integrin ITGA1/ITGB1 heterodimer. Interacts with host integrin ITGA2/ITGB1 heterodimer. Interaction with microtubules blocks trafficking to the Golgi apparatus. Ref.3 Ref.7 Ref.8 Ref.10 Ref.12 Ref.13 Ref.14

Subcellular location

Non-structural glycoprotein 4: Host rough endoplasmic reticulum membrane; Single-pass type III membrane protein By similarity. Host membranehost caveola; Single-pass type III membrane protein. Secreted By similarity. Note: Immature double-layered particles assembled in the cytoplasm bud across the membrane of the endoplasmic reticulum, acquiring during this process a transient lipid membrane that is modified with the ER resident viral glycoproteins NSP4 and VP7; these enveloped particles also contain VP4. As the particles move towards the interior of the ER cisternae, the transient lipid membrane and the non-structural protein NSP4 are lost, while the virus surface proteins VP4 and VP7 rearrange to form the outermost virus protein layer, yielding mature infectious triple-layered particles. NSP4 also localizes in vesicular structures, which contain an autophagosomal marker and associate with viroplasms in virus-infected cells. Ref.1 Ref.3 Ref.7 Ref.10 Ref.11 Ref.12

Domain

A disordered 28 AA C-terminal domain is presented to the cytoplasm by each subunit of the tetrameric receptor. Ref.5

The coiled coil region mediates oligomerization. Ref.5

Post-translational modification

The N-glycosyl content is primarily Man9GlcNAc, with a small amount of Man8GlcNAc.

Miscellaneous

Binds 1 calcium ion per tetramer. The calcium ion is bound by a glutamine from each tetrameric partner and by a glutamic acid from two of the tetrameric partners, while the glutamic acid from the other two partners do not participate in binding the ion By similarity.

Sequence similarities

Belongs to the rotavirus NSP4 family.

Caution

A candidate enterotoxigenic cleaved form of the protein has been suggested [Ref.2], but it remains unclear whether this truncated form constitutes an active enterotoxin in vivo.

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 175175Non-structural glycoprotein 4
PRO_0000149628

Regions

Topological domain1 – 2828Lumenal
Transmembrane29 – 5123Helical; Potential
Topological domain52 – 175124Cytoplasmic
Region7 – 2115Hydrophobic By similarity
Region67 – 8519Hydrophobic By similarity
Region85 – 12339Endoplasmic reticulum retention signal Potential
Region114 – 13522Interaction with CAV1 By similarity
Region122 – 17554Required for interaction with microtubules
Region159 – 17517ICP binding domain
Coiled coil85 – 11733 Potential

Sites

Metal binding1201Calcium; shared with all tetrameric partners; partial
Metal binding1231Calcium; shared with all tetrameric partners
Site111 – 1122Cleavage Potential

Amino acid modifications

Glycosylation81N-linked (GlcNAc...) (high mannose); by host Ref.1
Glycosylation181N-linked (GlcNAc...) (high mannose); by host Ref.1

Experimental info

Mutagenesis631C → S: No effect on receptor activity nor on oligomer formation. No effect on receptor activity nor on oligomer formation; when associated with C-71. Ref.3
Mutagenesis711C → S: No effect on receptor activity nor on oligomer formation. No effect on receptor activity nor on oligomer formation; when associated with C-63. Ref.3
Mutagenesis1201E → A: Complete loss of interaction with integrin ITGA2. Ref.13
Mutagenesis1661Y → I: Loss of ICP-binding activity. Ref.8
Mutagenesis174 – 1752Missing: Abolishes receptor activity. Ref.3
Mutagenesis1751M → G, K or I: Abolishes receptor activity. Ref.3
Mutagenesis1751Missing: Abolishes receptor activity. No effect on subcellular location. Ref.3

Secondary structure

... 175
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
P04512 [UniParc].

Last modified August 13, 1987. Version 1.
Checksum: 1E72A3223C567309

FASTA17520,267
        10         20         30         40         50         60 
MEKLTDLNYT LSVITLMNNT LHTILEDPGM AYFPYIASVL TGLFALNKAS IPTMKIALKT 

        70         80         90        100        110        120 
SKCSYKVVKY CIVTIFNTLL KLAGYKEQIT TKDEIEKQMD RVVKEMRRQL EMIDKLTTRE 

       130        140        150        160        170 
IEQVELLKRI YDKLTVQTTG EIDMTKEINQ KNVRTLEEWE SGKNPYEPRE VTAAM 

« Hide

References

[1]"Coding assignment and nucleotide sequence of simian rotavirus SA11 gene segment 10: location of glycosylation sites suggests that the signal peptide is not cleaved."
Both G.W., Siegman L.J., Bellamy A.R., Atkinson P.H.
J. Virol. 48:335-339(1983) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [GENOMIC RNA], SUBCELLULAR LOCATION, CARBOHYDRATE STRUCTURE, GLYCOSYLATION AT ASN-8 AND ASN-18.
[2]"A functional NSP4 enterotoxin peptide secreted from rotavirus-infected cells."
Zhang M., Zeng C.Q.-Y., Morris A.P., Estes M.K.
J. Virol. 74:11663-11670(2000) [PubMed] [Europe PMC] [Abstract]
Cited for: PROTEIN SEQUENCE OF 112-120, PROTEOLYTIC CLEAVAGE, FUNCTION.
[3]"Transient expression and mutational analysis of the rotavirus intracellular receptor: the C-terminal methionine residue is essential for ligand binding."
Taylor J.A., Meyer J.C., Legge M.A., O'Brien J.A., Street J.E., Lord V.J., Bergmann C.C., Bellamy A.R.
J. Virol. 66:3566-3572(1992) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION, SUBUNIT, SUBCELLULAR LOCATION, MUTAGENESIS OF CYS-63; CYS-71; MET-175 AND 174-ALA--MET-175.
[4]"The rotavirus nonstructural glycoprotein NSP4 mobilizes Ca2+ from the endoplasmic reticulum."
Tian P., Estes M.K., Hu Y., Ball J.M., Zeng C.Q.-Y., Schilling W.P.
J. Virol. 69:5763-5772(1995) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION.
[5]"The cytoplasmic tail of NSP4, the endoplasmic reticulum-localized non-structural glycoprotein of rotavirus, contains distinct virus binding and coiled coil domains."
Taylor J.A., O'Brien J.A., Yeager M.
EMBO J. 15:4469-4476(1996) [PubMed] [Europe PMC] [Abstract]
Cited for: COILED-COIL DOMAIN.
[6]"The rotavirus enterotoxin NSP4 mobilizes intracellular calcium in human intestinal cells by stimulating phospholipase C-mediated inositol 1,4,5-trisphosphate production."
Dong Y., Zeng C.Q.-Y., Ball J.M., Estes M.K., Morris A.P.
Proc. Natl. Acad. Sci. U.S.A. 94:3960-3965(1997) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION.
[7]"Immobilization of the early secretory pathway by a virus glycoprotein that binds to microtubules."
Xu A., Bellamy A.R., Taylor J.A.
EMBO J. 19:6465-6474(2000) [PubMed] [Europe PMC] [Abstract]
Cited for: INTERACTION WITH MICROTUBULES, SUBCELLULAR LOCATION.
[8]"Probing the structure of rotavirus NSP4: a short sequence at the extreme C terminus mediates binding to the inner capsid particle."
O'Brien J.A., Taylor J.A., Bellamy A.R.
J. Virol. 74:5388-5394(2000) [PubMed] [Europe PMC] [Abstract]
Cited for: INTERACTION WITH ICP, MUTAGENESIS OF TYR-166.
[9]"A cytoplasmic region of the NSP4 enterotoxin of rotavirus is involved in retention in the endoplasmic reticulum."
Mirazimi A., Magnusson K.-E., Svensson L.
J. Gen. Virol. 84:875-883(2003) [PubMed] [Europe PMC] [Abstract]
Cited for: ER RETENTION SIGNAL.
[10]"The rotavirus enterotoxin NSP4 directly interacts with the caveolar structural protein caveolin-1."
Parr R.D., Storey S.M., Mitchell D.M., McIntosh A.L., Zhou M., Mir K.D., Ball J.M.
J. Virol. 80:2842-2854(2006) [PubMed] [Europe PMC] [Abstract]
Cited for: INTERACTION WITH HUMAN CAV1, SUBCELLULAR LOCATION.
[11]"Rotavirus NSP4 induces a novel vesicular compartment regulated by calcium and associated with viroplasms."
Berkova Z., Crawford S.E., Trugnan G., Yoshimori T., Morris A.P., Estes M.K.
J. Virol. 80:6061-6071(2006) [PubMed] [Europe PMC] [Abstract]
Cited for: SUBCELLULAR LOCATION.
[12]"Full-length, glycosylated NSP4 is localized to plasma membrane caveolae by a novel raft isolation technique."
Storey S.M., Gibbons T.F., Williams C.V., Parr R.D., Schroeder F., Ball J.M.
J. Virol. 81:5472-5483(2007) [PubMed] [Europe PMC] [Abstract]
Cited for: INTERACTION WITH HUMAN CAV1, SUBCELLULAR LOCATION, GLYCOSYLATION.
Strain: SA11-4F.
[13]"Inaugural article: integrins alpha1beta1 and alpha2beta1 are receptors for the rotavirus enterotoxin."
Seo N.-S., Zeng C.Q.-Y., Hyser J.M., Utama B., Crawford S.E., Kim K.J., Hoeoek M., Estes M.K.
Proc. Natl. Acad. Sci. U.S.A. 105:8811-8818(2008) [PubMed] [Europe PMC] [Abstract]
Cited for: INTERACTION WITH HUMAN INTEGRIN ITGA1/ITGB1 HETERODIMER, INTERACTION WITH HUMAN INTEGRIN ITGA2/ITGB1 HETERODIMER, MUTAGENESIS OF GLU-120.
[14]"Crystal structure of the oligomerization domain of NSP4 from rotavirus reveals a core metal-binding site."
Bowman G.D., Nodelman I.M., Levy O., Lin S.L., Tian P., Zamb T.J., Udem S.A., Venkataraghavan B., Schutt C.E.
J. Mol. Biol. 304:861-871(2000) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.86 ANGSTROMS) OF 95-137 IN COMPLEX WITH CALCIUM OR STRONTIUM, METAL-BINDING AT GLN-123 AND GLU-120, SUBUNIT.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
K01138 Genomic RNA. Translation: AAA47291.1.
PIRVGXRTS. A04140.

3D structure databases

PDBe
RCSB PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
1G1IX-ray2.00A/B95-137[»]
1G1JX-ray1.86A/B95-137[»]
2O1KX-ray1.67A/B95-146[»]
ProteinModelPortalP04512.
SMRP04512. Positions 95-137.
ModBaseSearch...
MobiDBSearch...

Protein-protein interaction databases

DIPDIP-46136N.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Family and domain databases

InterProIPR002107. Rotavirus_NSP4.
[Graphical view]
PfamPF01452. Rota_NSP4. 1 hit.
[Graphical view]
ProDomPD002202. NSP4_rotavirus. 1 hit.
[Graphical view] [Entries sharing at least one domain]
ProtoNetSearch...

Other

EvolutionaryTraceP04512.

Entry information

Entry nameNSP4_ROTS1
AccessionPrimary (citable) accession number: P04512
Entry history
Integrated into UniProtKB/Swiss-Prot: August 13, 1987
Last sequence update: August 13, 1987
Last modified: February 19, 2014
This is version 83 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programViral Protein Annotation Program

Relevant documents

SIMILARITY comments

Index of protein domains and families

PDB cross-references

Index of Protein Data Bank (PDB) cross-references