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==C425S mutant of the C-terminal cytoplasmic domain of non-structural protein 4 from mouse hepatitis virus A59== | |||
<StructureSection load='3vcb' size='340' side='right' caption='[[3vcb]], [[Resolution|resolution]] 2.40Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[3vcb]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Murine_coronavirus_inf-mhv-a59 Murine coronavirus inf-mhv-a59]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3VCB OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3VCB FirstGlance]. <br> | |||
</td></tr><tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">Murine hepatitis virus strain A59 ORF1ab, ORF1A ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=591071 Murine coronavirus inf-MHV-A59])</td></tr> | |||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3vcb FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3vcb OCA], [http://www.rcsb.org/pdb/explore.do?structureId=3vcb RCSB], [http://www.ebi.ac.uk/pdbsum/3vcb PDBsum]</span></td></tr> | |||
</table> | |||
== Function == | |||
[[http://www.uniprot.org/uniprot/Q9J3E9_9BETC Q9J3E9_9BETC]] Nsp7-nsp8 hexadecamer may possibly confer processivity to the polymerase, maybe by binding to dsRNA or by producing primers utilized by the latter (By similarity).[SAAS:SAAS014828_004_014545] Nsp9 is a ssRNA-binding protein (By similarity).[SAAS:SAAS014828_004_001449] | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
BACKGROUND: The replication of coronaviruses takes place on cytoplasmic double membrane vesicles (DMVs) originating in the endoplasmic reticulum (ER). Three trans-membrane non-structural proteins, nsp3, nsp4 and nsp6, are understood to be membrane anchors of the coronavirus replication complex. Nsp4 is localized to the ER membrane when expressed alone but is recruited into the replication complex in infected cells. It is revealed to contain four trans-membrane regions and its N- and C-termini are exposed to the cytosol. METHODOLOGY/PRINCIPAL FINDINGS: We have determined the crystal structures of the C-terminal hydrophilic domain of nsp4 (nsp4C) from MHV strain A59 and a C425S site-directed mutant. The highly conserved 89 amino acid region from T408 to Q496 is shown to possess a new fold. The wild-type (WT) structure features two monomers linked by a Cys425-Cys425 disulfide bond in one asymmetric unit. The monomers are arranged with their N- and C-termini in opposite orientations to form an "open" conformation. Mutation of Cys425 to Ser did not affect the monomer structure, although the mutant dimer adopts strikingly different conformations by crystal packing, with the cross-linked C-termini and parallel N-termini of two monomers forming a "closed" conformation. The WT nsp4C exists as a dimer in solution and can dissociate easily into monomers in a reducing environment. CONCLUSIONS/SIGNIFICANCE: As nsp4C is exposed in the reducing cytosol, the monomer of nsp4C should be physiological. This structure may serve as a basis for further functional studies of nsp4. | |||
Crystal structure of the C-terminal cytoplasmic domain of non-structural protein 4 from mouse hepatitis virus A59.,Xu X, Lou Z, Ma Y, Chen X, Yang Z, Tong X, Zhao Q, Xu Y, Deng H, Bartlam M, Rao Z PLoS One. 2009 Jul 10;4(7):e6217. PMID:19593433<ref>PMID:19593433</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
==See Also== | ==See Also== | ||
*[[RNA polymerase|RNA polymerase]] | *[[RNA polymerase|RNA polymerase]] | ||
== References == | |||
== | <references/> | ||
__TOC__ | |||
</StructureSection> | |||
[[Category: Murine coronavirus inf-mhv-a59]] | [[Category: Murine coronavirus inf-mhv-a59]] | ||
[[Category: Bartlam, M | [[Category: Bartlam, M]] | ||
[[Category: Chen, X | [[Category: Chen, X]] | ||
[[Category: Deng, H | [[Category: Deng, H]] | ||
[[Category: Lou, Z | [[Category: Lou, Z]] | ||
[[Category: Ma, Y | [[Category: Ma, Y]] | ||
[[Category: Rao, Z | [[Category: Rao, Z]] | ||
[[Category: Tong, X | [[Category: Tong, X]] | ||
[[Category: Xu, X | [[Category: Xu, X]] | ||
[[Category: Xu, Y | [[Category: Xu, Y]] | ||
[[Category: Yang, Z | [[Category: Yang, Z]] | ||
[[Category: Zhao, Q | [[Category: Zhao, Q]] | ||
[[Category: Cytoplasmic]] | [[Category: Cytoplasmic]] | ||
[[Category: Homodimer]] | [[Category: Homodimer]] | ||
Revision as of 23:44, 24 December 2014
C425S mutant of the C-terminal cytoplasmic domain of non-structural protein 4 from mouse hepatitis virus A59C425S mutant of the C-terminal cytoplasmic domain of non-structural protein 4 from mouse hepatitis virus A59
Structural highlights
Function[Q9J3E9_9BETC] Nsp7-nsp8 hexadecamer may possibly confer processivity to the polymerase, maybe by binding to dsRNA or by producing primers utilized by the latter (By similarity).[SAAS:SAAS014828_004_014545] Nsp9 is a ssRNA-binding protein (By similarity).[SAAS:SAAS014828_004_001449] Publication Abstract from PubMedBACKGROUND: The replication of coronaviruses takes place on cytoplasmic double membrane vesicles (DMVs) originating in the endoplasmic reticulum (ER). Three trans-membrane non-structural proteins, nsp3, nsp4 and nsp6, are understood to be membrane anchors of the coronavirus replication complex. Nsp4 is localized to the ER membrane when expressed alone but is recruited into the replication complex in infected cells. It is revealed to contain four trans-membrane regions and its N- and C-termini are exposed to the cytosol. METHODOLOGY/PRINCIPAL FINDINGS: We have determined the crystal structures of the C-terminal hydrophilic domain of nsp4 (nsp4C) from MHV strain A59 and a C425S site-directed mutant. The highly conserved 89 amino acid region from T408 to Q496 is shown to possess a new fold. The wild-type (WT) structure features two monomers linked by a Cys425-Cys425 disulfide bond in one asymmetric unit. The monomers are arranged with their N- and C-termini in opposite orientations to form an "open" conformation. Mutation of Cys425 to Ser did not affect the monomer structure, although the mutant dimer adopts strikingly different conformations by crystal packing, with the cross-linked C-termini and parallel N-termini of two monomers forming a "closed" conformation. The WT nsp4C exists as a dimer in solution and can dissociate easily into monomers in a reducing environment. CONCLUSIONS/SIGNIFICANCE: As nsp4C is exposed in the reducing cytosol, the monomer of nsp4C should be physiological. This structure may serve as a basis for further functional studies of nsp4. Crystal structure of the C-terminal cytoplasmic domain of non-structural protein 4 from mouse hepatitis virus A59.,Xu X, Lou Z, Ma Y, Chen X, Yang Z, Tong X, Zhao Q, Xu Y, Deng H, Bartlam M, Rao Z PLoS One. 2009 Jul 10;4(7):e6217. PMID:19593433[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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