ID   VP4_ROTHW               Reviewed;         775 AA.
AC   P11193; Q05334; Q86202;
DT   01-JUL-1989, integrated into UniProtKB/Swiss-Prot.
DT   14-APR-2009, sequence version 3.
DT   24-JAN-2024, entry version 138.
DE   RecName: Full=Outer capsid protein VP4 {ECO:0000255|HAMAP-Rule:MF_04132};
DE   AltName: Full=Hemagglutinin {ECO:0000255|HAMAP-Rule:MF_04132};
DE   Contains:
DE     RecName: Full=Outer capsid protein VP8* {ECO:0000255|HAMAP-Rule:MF_04132};
DE   Contains:
DE     RecName: Full=Outer capsid protein VP5* {ECO:0000255|HAMAP-Rule:MF_04132};
OS   Rotavirus A (strain RVA/Human/United States/Wa/1974/G1P1A[8]) (RV-A).
OC   Viruses; Riboviria; Orthornavirae; Duplornaviricota; Resentoviricetes;
OC   Reovirales; Sedoreoviridae; Rotavirus; Rotavirus A.
OX   NCBI_TaxID=10962;
OH   NCBI_TaxID=9606; Homo sapiens (Human).
RN   [1]
RP   NUCLEOTIDE SEQUENCE.
RX   PubMed=2839714; DOI=10.1128/jvi.62.8.2978-2984.1988;
RA   Gorziglia M., Green K.Y., Nishikawa K., Taniguchi K., Jones R.W.,
RA   Kapikian A.Z., Chanock R.M.;
RT   "Sequence of the fourth gene of human rotaviruses recovered from
RT   asymptomatic or symptomatic infections.";
RL   J. Virol. 62:2978-2984(1988).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [GENOMIC RNA].
RX   PubMed=8389116; DOI=10.1007/bf01319006;
RA   Kitamoto N., Mattion N.M., Estes M.K.;
RT   "Alterations in the sequence of the gene 4 from a human rotavirus after
RT   multiple passages in HepG2 liver cells.";
RL   Arch. Virol. 130:179-185(1993).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [GENOMIC RNA].
RX   PubMed=7530390; DOI=10.1016/s0042-6822(95)80029-8;
RA   Padilla-Noriega L., Dunn S.J., Lopez S., Greenberg H.B., Arias C.F.;
RT   "Identification of two independent neutralization domains on the VP4
RT   trypsin cleavage products VP5* and VP8* of human rotavirus ST3.";
RL   Virology 206:148-154(1995).
RN   [4]
RP   NUCLEOTIDE SEQUENCE [GENOMIC RNA] OF 1-280.
RX   PubMed=3018754; DOI=10.1073/pnas.83.18.7039;
RA   Gorziglia M., Hoshino Y., Buckler-White A., Blumentals I., Glass R.,
RA   Flores J., Kapikian A.Z., Chanock R.M.;
RT   "Conservation of amino acid sequence of VP8 and cleavage region of 84-kDa
RT   outer capsid protein among rotaviruses recovered from asymptomatic neonatal
RT   infection.";
RL   Proc. Natl. Acad. Sci. U.S.A. 83:7039-7043(1986).
RN   [5]
RP   INTERACTION WITH HUMAN INTEGRIN HETERODIMER ITGA4/ITGB1 (OUTER CAPSID
RP   VP5*), AND INTERACTION WITH HUMAN INTEGRIN HETERODIMER ITGA4/ITGB7 (OUTER
RP   CAPSID PROTEIN VP5*).
RX   PubMed=16298987; DOI=10.1099/vir.0.81102-0;
RA   Graham K.L., Fleming F.E., Halasz P., Hewish M.J., Nagesha H.S.,
RA   Holmes I.H., Takada Y., Coulson B.S.;
RT   "Rotaviruses interact with alpha4beta7 and alpha4beta1 integrins by binding
RT   the same integrin domains as natural ligands.";
RL   J. Gen. Virol. 86:3397-3408(2005).
RN   [6]
RP   INTERACTION WITH HUMAN INTEGRIN HETERODIMER ITGA2/ITGB1 (OUTER CAPSID
RP   PROTEIN VP5*).
RX   PubMed=16603530; DOI=10.1099/vir.0.81580-0;
RA   Graham K.L., Takada Y., Coulson B.S.;
RT   "Rotavirus spike protein VP5* binds alpha2beta1 integrin on the cell
RT   surface and competes with virus for cell binding and infectivity.";
RL   J. Gen. Virol. 87:1275-1283(2006).
RN   [7]
RP   REVIEW.
RX   PubMed=16575520; DOI=10.1007/s10719-006-5435-y;
RA   Isa P., Arias C.F., Lopez S.;
RT   "Role of sialic acids in rotavirus infection.";
RL   Glycoconj. J. 23:27-37(2006).
RN   [8]
RP   FUNCTION (OUTER CAPSID PROTEIN VP8*).
RX   PubMed=24501414; DOI=10.1128/jvi.03431-13;
RA   Fleming F.E., Boehm R., Dang V.T., Holloway G., Haselhorst T., Madge P.D.,
RA   Deveryshetty J., Yu X., Blanchard H., von Itzstein M., Coulson B.S.;
RT   "Relative roles of GM1 ganglioside, N-acylneuraminic acids, and alpha2beta1
RT   integrin in mediating rotavirus infection.";
RL   J. Virol. 88:4558-4571(2014).
RN   [9]
RP   X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) OF 64-223, AND FUNCTION (OUTER CAPSID
RP   PROTEIN VP8*).
RX   PubMed=17306299; DOI=10.1016/j.jmb.2007.01.028;
RA   Blanchard H., Yu X., Coulson B.S., von Itzstein M.;
RT   "Insight into host cell carbohydrate-recognition by human and porcine
RT   rotavirus from crystal structures of the virion spike associated
RT   carbohydrate-binding domain (VP8*).";
RL   J. Mol. Biol. 367:1215-1226(2007).
CC   -!- FUNCTION: [Outer capsid protein VP4]: Spike-forming protein that
CC       mediates virion attachment to the host epithelial cell receptors and
CC       plays a major role in cell penetration, determination of host range
CC       restriction and virulence. Rotavirus attachment and entry into the host
CC       cell probably involves multiple sequential contacts between the outer
CC       capsid proteins VP4 and VP7, and the cell receptors. It is subsequently
CC       lost, together with VP7, following virus entry into the host cell.
CC       Following entry into the host cell, low intracellular or intravesicular
CC       Ca(2+) concentration probably causes the calcium-stabilized VP7 trimers
CC       to dissociate from the virion. This step is probably necessary for the
CC       membrane-disrupting entry step and the release of VP4, which is locked
CC       onto the virion by VP7. During the virus exit from the host cell, VP4
CC       seems to be required to target the newly formed virions to the host
CC       cell lipid rafts. {ECO:0000255|HAMAP-Rule:MF_04132}.
CC   -!- FUNCTION: [Outer capsid protein VP5*]: Forms the spike 'foot' and
CC       'body' and acts as a membrane permeabilization protein that mediates
CC       release of viral particles from endosomal compartments into the
CC       cytoplasm. During entry, the part of VP5* that protrudes from the virus
CC       folds back on itself and reorganizes from a local dimer to a trimer.
CC       This reorganization may be linked to membrane penetration by exposing
CC       VP5* hydrophobic region. In integrin-dependent strains, VP5* targets
CC       the integrin heterodimer ITGA2/ITGB1 for cell attachment.
CC       {ECO:0000255|HAMAP-Rule:MF_04132}.
CC   -!- FUNCTION: VP8* Forms the head of the spikes and mediates the
CC       recognition of specific host cell surface glycans (PubMed:17306299). It
CC       is the viral hemagglutinin and an important target of neutralizing
CC       antibodies (By similarity). In sialic acid-dependent strains, VP8*
CC       binds to host cell sialic acid, most probably a ganglioside, providing
CC       the initial contact (PubMed:24501414). In some other strains, VP8*
CC       mediates the attachment to histo-blood group antigens (HBGAs) for viral
CC       entry (By similarity). {ECO:0000255|HAMAP-Rule:MF_04132,
CC       ECO:0000269|PubMed:17306299, ECO:0000269|PubMed:24501414}.
CC   -!- SUBUNIT: [Outer capsid protein VP4]: Homotrimer. VP4 adopts a dimeric
CC       appearance above the capsid surface, while forming a trimeric base
CC       anchored inside the capsid layer. Only hints of the third molecule are
CC       observed above the capsid surface. It probably performs a series of
CC       molecular rearrangements during viral entry. Prior to trypsin cleavage,
CC       it is flexible. The priming trypsin cleavage triggers its rearrangement
CC       into rigid spikes with approximate two-fold symmetry of their
CC       protruding parts. After an unknown second triggering event, cleaved VP4
CC       may undergo another rearrangement, in which two VP5* subunits fold back
CC       on themselves and join a third subunit to form a tightly associated
CC       trimer, shaped like a folded umbrella. Interacts with VP6. Interacts
CC       with VP7. {ECO:0000255|HAMAP-Rule:MF_04132}.
CC   -!- SUBUNIT: [Outer capsid protein VP5*]: Homotrimer. The trimer is coiled-
CC       coil stabilized by its C-terminus, however, its N-terminus, known as
CC       antigen domain or 'body', seems to be flexible allowing it to self-
CC       associate either as a dimer or a trimer (By similarity). Interacts with
CC       host ITGA2 (via ITAG2 I-domain); this interaction occurs when ITGA2 is
CC       part of the integrin heterodimer ITGA2/ITGB1 (PubMed:16603530).
CC       Interacts with host integrin heterodimer ITGA4/ITGB1 and ITGA4/ITGB7
CC       (PubMed:16298987). {ECO:0000255|HAMAP-Rule:MF_04132,
CC       ECO:0000269|PubMed:16298987, ECO:0000269|PubMed:16603530}.
CC   -!- SUBCELLULAR LOCATION: [Outer capsid protein VP4]: Virion
CC       {ECO:0000255|HAMAP-Rule:MF_04132}. Host rough endoplasmic reticulum
CC       {ECO:0000255|HAMAP-Rule:MF_04132}. Host cell membrane
CC       {ECO:0000255|HAMAP-Rule:MF_04132}. Host cytoplasm, host cytoskeleton
CC       {ECO:0000255|HAMAP-Rule:MF_04132}. Host endoplasmic reticulum-Golgi
CC       intermediate compartment {ECO:0000255|HAMAP-Rule:MF_04132}. Note=The
CC       outer layer contains 180 copies of VP4, grouped as 60 dimers. Immature
CC       double-layered particles assembled in the cytoplasm bud across the
CC       membrane of the endoplasmic reticulum, acquiring during this process a
CC       transient lipid membrane that is modified with the ER resident viral
CC       glycoproteins NSP4 and VP7; these enveloped particles also contain VP4.
CC       As the particles move towards the interior of the ER cisternae, the
CC       transient lipid membrane and the non-structural protein NSP4 are lost,
CC       while the virus surface proteins VP4 and VP7 rearrange to form the
CC       outermost virus protein layer, yielding mature infectious triple-
CC       layered particles. VP4 also seems to associate with lipid rafts of the
CC       host cell membrane probably for the exit of the virus from the infected
CC       cell by an alternate pathway. {ECO:0000255|HAMAP-Rule:MF_04132}.
CC   -!- SUBCELLULAR LOCATION: [Outer capsid protein VP8*]: Virion
CC       {ECO:0000255|HAMAP-Rule:MF_04132}. Note=Outer capsid protein.
CC       {ECO:0000255|HAMAP-Rule:MF_04132}.
CC   -!- SUBCELLULAR LOCATION: [Outer capsid protein VP5*]: Virion
CC       {ECO:0000255|HAMAP-Rule:MF_04132}. Note=Outer capsid protein.
CC       {ECO:0000255|HAMAP-Rule:MF_04132}.
CC   -!- DOMAIN: [Outer capsid protein VP4]: The VP4 spike is divided into a
CC       foot, a stalk and body, and a head. {ECO:0000255|HAMAP-Rule:MF_04132}.
CC   -!- PTM: [Outer capsid protein VP4]: Proteolytic cleavage by trypsin
CC       results in activation of VP4 functions and greatly increases
CC       infectivity. The penetration into the host cell is dependent on trypsin
CC       treatment of VP4. It produces two peptides, VP5* and VP8* that remain
CC       associated with the virion. Cleavage of VP4 by trypsin probably occurs
CC       in vivo in the lumen of the intestine prior to infection of
CC       enterocytes. Trypsin seems to be incorporated into the three-layered
CC       viral particles but remains inactive as long as the viral outer capsid
CC       is intact and would only be activated upon the solubilization of the
CC       latter. {ECO:0000255|HAMAP-Rule:MF_04132}.
CC   -!- MISCELLANEOUS: This strain probably does not use sialic acid to attach
CC       to the host cell. {ECO:0000303|PubMed:16575520}.
CC   -!- MISCELLANEOUS: In group A rotaviruses, VP4 defines the P serotype.
CC       {ECO:0000255|HAMAP-Rule:MF_04132}.
CC   -!- MISCELLANEOUS: Some rotavirus strains are neuraminidase-sensitive and
CC       require sialic acid to attach to the cell surface. Some rotavirus
CC       strains are integrin-dependent. Some rotavirus strains depend on
CC       ganglioside for their entry into the host cell. Hsp70 also seems to be
CC       involved in the entry of some strains. {ECO:0000255|HAMAP-
CC       Rule:MF_04132}.
CC   -!- SIMILARITY: Belongs to the rotavirus VP4 family. {ECO:0000255|HAMAP-
CC       Rule:MF_04132}.
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DR   EMBL; M96825; AAA47290.1; -; Genomic_RNA.
DR   EMBL; L34161; AAA66953.1; -; Genomic_RNA.
DR   PIR; A28839; VPXRW3.
DR   PDB; 2DWR; X-ray; 2.50 A; A=65-223.
DR   PDBsum; 2DWR; -.
DR   SMR; P11193; -.
DR   UniLectin; P11193; -.
DR   ABCD; P11193; 18 sequenced antibodies.
DR   EvolutionaryTrace; P11193; -.
DR   Proteomes; UP000006581; Genome.
DR   GO; GO:0044172; C:host cell endoplasmic reticulum-Golgi intermediate compartment; IEA:UniProtKB-SubCell.
DR   GO; GO:0020002; C:host cell plasma membrane; IEA:UniProtKB-SubCell.
DR   GO; GO:0044168; C:host cell rough endoplasmic reticulum; IEA:UniProtKB-SubCell.
DR   GO; GO:0044163; C:host cytoskeleton; IEA:UniProtKB-SubCell.
DR   GO; GO:0016020; C:membrane; IEA:UniProtKB-KW.
DR   GO; GO:0039624; C:viral outer capsid; IEA:UniProtKB-UniRule.
DR   GO; GO:0039665; P:permeabilization of host organelle membrane involved in viral entry into host cell; IEA:UniProtKB-UniRule.
DR   GO; GO:0099008; P:viral entry via permeabilization of inner membrane; IEA:UniProtKB-KW.
DR   GO; GO:0019062; P:virion attachment to host cell; IEA:UniProtKB-UniRule.
DR   Gene3D; 1.20.5.170; -; 1.
DR   Gene3D; 2.60.120.200; -; 1.
DR   HAMAP; MF_04132; Rota_A_VP4; 1.
DR   HAMAP; MF_04125; Rota_VP4; 1.
DR   InterPro; IPR013320; ConA-like_dom_sf.
DR   InterPro; IPR042546; Rota_A_VP4.
DR   InterPro; IPR035330; Rota_VP4_MID.
DR   InterPro; IPR038017; Rota_VP4_MID_sf.
DR   InterPro; IPR000416; VP4_concanavalin-like.
DR   InterPro; IPR035329; VP4_helical.
DR   Pfam; PF17477; Rota_VP4_MID; 1.
DR   Pfam; PF00426; VP4_haemagglut; 1.
DR   Pfam; PF17478; VP4_helical; 1.
DR   SUPFAM; SSF49899; Concanavalin A-like lectins/glucanases; 1.
DR   SUPFAM; SSF111379; VP4 membrane interaction domain; 1.
PE   1: Evidence at protein level;
KW   3D-structure; Capsid protein; Coiled coil; Disulfide bond; Hemagglutinin;
KW   Host cell membrane; Host cytoplasm; Host cytoskeleton;
KW   Host endoplasmic reticulum; Host membrane; Host-virus interaction;
KW   Membrane; Outer capsid protein; Viral attachment to host cell;
KW   Viral penetration into host cytoplasm;
KW   Viral penetration via permeabilization of host membrane; Virion;
KW   Virus entry into host cell.
FT   CHAIN           1..775
FT                   /note="Outer capsid protein VP4"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_04132"
FT                   /id="PRO_0000041090"
FT   CHAIN           1..230
FT                   /note="Outer capsid protein VP8*"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_04132"
FT                   /id="PRO_0000041091"
FT   CHAIN           247..775
FT                   /note="Outer capsid protein VP5*"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_04132"
FT                   /id="PRO_0000041092"
FT   REGION          65..223
FT                   /note="Spike head"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_04132"
FT   REGION          247..479
FT                   /note="Antigen domain"
FT   REGION          247..478
FT                   /note="Spike body and stalk (antigen domain)"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_04132"
FT   REGION          307..309
FT                   /note="DGE motif; interaction with ITGA2/ITGB1 heterodimer"
FT                   /evidence="ECO:0000269|PubMed:16603530"
FT   REGION          388..408
FT                   /note="Hydrophobic; possible role in virus entry into host
FT                   cell"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_04132"
FT   REGION          509..775
FT                   /note="Spike foot"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_04132"
FT   COILED          483..517
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_04132"
FT   MOTIF           307..309
FT                   /note="DGE motif; interaction with ITGA2/ITGB1 heterodimer"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_04132"
FT   MOTIF           447..449
FT                   /note="YGL motif; interaction with ITGA4"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_04132"
FT   MOTIF           643..645
FT                   /note="KID motif; interaction with HSPA8"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_04132"
FT   SITE            230..231
FT                   /note="Cleavage"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_04132"
FT   SITE            240..241
FT                   /note="Cleavage"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_04132"
FT   SITE            246..247
FT                   /note="Cleavage; associated with enhancement of
FT                   infectivity"
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_04132"
FT   DISULFID        317..379
FT                   /evidence="ECO:0000255|HAMAP-Rule:MF_04132"
FT   CONFLICT        64
FT                   /note="I -> M (in Ref. 3; AAA66953)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        100
FT                   /note="D -> E (in Ref. 4)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        120
FT                   /note="T -> L (in Ref. 1)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        265
FT                   /note="Q -> E (in Ref. 4)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        331
FT                   /note="S -> F (in Ref. 3; AAA66953)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        388
FT                   /note="I -> L (in Ref. 2; AAA47290)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        525
FT                   /note="D -> G (in Ref. 2; AAA47290)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        618
FT                   /note="F -> L (in Ref. 1)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        749
FT                   /note="N -> K (in Ref. 1)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        759
FT                   /note="N -> H (in Ref. 1)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        774
FT                   /note="K -> R (in Ref. 1)"
FT                   /evidence="ECO:0000305"
FT   STRAND          66..69
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   STRAND          71..74
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   STRAND          80..85
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   STRAND          88..96
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   STRAND          98..100
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   STRAND          102..108
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   STRAND          110..121
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   STRAND          124..132
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   STRAND          134..146
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   STRAND          152..161
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   STRAND          163..169
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   STRAND          172..179
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   STRAND          184..189
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   STRAND          197..200
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   STRAND          204..208
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   HELIX           209..211
FT                   /evidence="ECO:0007829|PDB:2DWR"
FT   HELIX           212..221
FT                   /evidence="ECO:0007829|PDB:2DWR"
SQ   SEQUENCE   775 AA;  87697 MW;  1CE181B859F74FB0 CRC64;
     MASLIYRQLL TNSYSVDLHD EIEQIGSEKT QNVTINPSPF AQTRYAPVNW GHGEINDSTT
     VEPILDGPYQ PTTFTPPNDY WILINSNTNG VVYESTNNSD FWTAVVAIEP HVNPVDRQYT
     IFGESKQFNV SNDSNKWKFL EMFRSSSQNE FYNRRTLTSD TRFVGILKYG GRVWTFHGET
     PRATTDSSST ANLNNISITI HSEFYIIPRS QESKCNEYIN NGLPPIQNTR NVVPLPLSSR
     SIQYKRAQVN EDIIVSKTSL WKEMQYNRDI IIRFKFGNSI VKMGGLGYKW SEISYKAANY
     QYNYLRDGEQ VTAHTTCSVN GVNNFSYNGG SLPTDFGISR YEVIKENSYV YVDYWDDSKA
     FRNMVYVRSL AANLNSVKCT GGSYNFSIPV GAWPVMNGGA VSLHFAGVTL STQFTDFVSL
     NSLRFRFSLT VDEPPFSILR TRTVNLYGLP AANPNNGNEY YEISGRFSLI YLVPTNDDYQ
     TPIMNSVTVR QDLERQLTDL REEFNSLSQE IAMAQLIDLA LLPLDMFSMF SGIKSTIDLT
     KSMATSVMKK FRKSKLATSI SEMTNSLSDA ASSASRNVSI RSNLSAISNW TNVSNDVSNV
     TNSLNDISTQ TSTISKKFRL KEMITQTEGM SFDDISAAVL KTKIDMSTQI GKNTLPDIVT
     EASEKFIPKR SYRILKDDEV MEINTEGKFF AYKINTFDEV PFDVNKFAEL VTDSPVISAI
     IDFKTLKNLN DNYGITRTEA LNLIKSNPNM LRNFINQNNP IIRNRIEQLI LQCKL
//