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==C425S mutant of the C-terminal cytoplasmic domain of non-structural protein 4 from mouse hepatitis virus A59==
==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]] 2.40&Aring;' scene=''>
<StructureSection load='3vcb' size='340' side='right'caption='[[3vcb]], [[Resolution|resolution]] 2.40&Aring;' scene=''>
== Structural highlights ==
== 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>
<table><tr><td colspan='2'>[[3vcb]] is a 2 chain structure with sequence from [https://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 [https://proteopedia.org/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>
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.4&#8491;</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://pdbe.org/3vcb PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=3vcb RCSB], [http://www.ebi.ac.uk/pdbsum/3vcb PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=3vcb ProSAT]</span></td></tr>
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=3vcb FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3vcb OCA], [https://pdbe.org/3vcb PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3vcb RCSB], [https://www.ebi.ac.uk/pdbsum/3vcb PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3vcb ProSAT]</span></td></tr>
</table>
</table>
== Function ==
== 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]
[https://www.uniprot.org/uniprot/R1AB_CVMA5 R1AB_CVMA5] The replicase polyprotein of coronaviruses is a multifunctional protein: it contains the activities necessary for the transcription of negative stranded RNA, leader RNA, subgenomic mRNAs and progeny virion RNA as well as proteinases responsible for the cleavage of the polyprotein into functional productsThe papain-like proteinase 1 (PL1-PRO) and papain-like proteinase 2 (PL2-PRO) are responsible for the cleavages located at the N-terminus of the replicase polyprotein. In addition, PLP2 possesses a deubiquitinating/deISGylating activity and processes both 'Lys-48'- and 'Lys-63'-linked polyubiquitin chains from cellular substrates. Antagonizes innate immune induction of type I interferon by blocking the phosphorylation, dimerization and subsequent nuclear translocation of host IRF-3 (By similarity). The main proteinase 3CL-PRO is responsible for the majority of cleavages as it cleaves the C-terminus of replicase polyprotein at 11 sites. Recognizes substrates containing the core sequence [ILMVF]-Q-|-[SGACN]. Inhibited by the substrate-analog Cbz-Val-Asn-Ser-Thr-Leu-Gln-CMK. Also contains an ADP-ribose-1''-phosphate (ADRP)-binding function (By similarity). The helicase which contains a zinc finger structure displays RNA and DNA duplex-unwinding activities with 5' to 3' polarity. ATPase activity is strongly stimulated by poly(U), poly(dT), poly(C), poly(dA), but not by poly(G) (By similarity). The exoribonuclease acts on both ssRNA and dsRNA in a 3' to 5' direction (By similarity). 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). Nsp9 is a ssRNA-binding protein (By similarity). NendoU is a Mn(2+)-dependent, uridylate-specific enzyme, which leaves 2'-3'-cyclic phosphates 5' to the cleaved bond (By similarity).
<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>
==See Also==
 
*[[RNA polymerase 3D structures|RNA polymerase 3D structures]]
From MEDLINE&reg;/PubMed&reg;, a database of the U.S. National Library of Medicine.<br>
</div>
<div class="pdbe-citations 3vcb" style="background-color:#fffaf0;"></div>
== References ==
<references/>
__TOC__
__TOC__
</StructureSection>
</StructureSection>
[[Category: Murine coronavirus inf-mhv-a59]]
[[Category: Large Structures]]
[[Category: Bartlam, M]]
[[Category: Murine coronavirus inf-MHV-A59]]
[[Category: Chen, X]]
[[Category: Bartlam M]]
[[Category: Deng, H]]
[[Category: Chen X]]
[[Category: Lou, Z]]
[[Category: Deng H]]
[[Category: Ma, Y]]
[[Category: Lou Z]]
[[Category: Rao, Z]]
[[Category: Ma Y]]
[[Category: Tong, X]]
[[Category: Rao Z]]
[[Category: Xu, X]]
[[Category: Tong X]]
[[Category: Xu, Y]]
[[Category: Xu X]]
[[Category: Yang, Z]]
[[Category: Xu Y]]
[[Category: Zhao, Q]]
[[Category: Yang Z]]
[[Category: Cytoplasmic]]
[[Category: Zhao Q]]
[[Category: Homodimer]]
[[Category: Host membrane]]
[[Category: Hydrolase]]
[[Category: Multi-pass membrane protein]]
[[Category: New fold]]
[[Category: Viral protein]]

Latest revision as of 13:35, 1 March 2024

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

3vcb is a 2 chain structure with sequence from Murine coronavirus inf-MHV-A59. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Method:X-ray diffraction, Resolution 2.4Å
Resources:FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

R1AB_CVMA5 The replicase polyprotein of coronaviruses is a multifunctional protein: it contains the activities necessary for the transcription of negative stranded RNA, leader RNA, subgenomic mRNAs and progeny virion RNA as well as proteinases responsible for the cleavage of the polyprotein into functional products. The papain-like proteinase 1 (PL1-PRO) and papain-like proteinase 2 (PL2-PRO) are responsible for the cleavages located at the N-terminus of the replicase polyprotein. In addition, PLP2 possesses a deubiquitinating/deISGylating activity and processes both 'Lys-48'- and 'Lys-63'-linked polyubiquitin chains from cellular substrates. Antagonizes innate immune induction of type I interferon by blocking the phosphorylation, dimerization and subsequent nuclear translocation of host IRF-3 (By similarity). The main proteinase 3CL-PRO is responsible for the majority of cleavages as it cleaves the C-terminus of replicase polyprotein at 11 sites. Recognizes substrates containing the core sequence [ILMVF]-Q-|-[SGACN]. Inhibited by the substrate-analog Cbz-Val-Asn-Ser-Thr-Leu-Gln-CMK. Also contains an ADP-ribose-1-phosphate (ADRP)-binding function (By similarity). The helicase which contains a zinc finger structure displays RNA and DNA duplex-unwinding activities with 5' to 3' polarity. ATPase activity is strongly stimulated by poly(U), poly(dT), poly(C), poly(dA), but not by poly(G) (By similarity). The exoribonuclease acts on both ssRNA and dsRNA in a 3' to 5' direction (By similarity). 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). Nsp9 is a ssRNA-binding protein (By similarity). NendoU is a Mn(2+)-dependent, uridylate-specific enzyme, which leaves 2'-3'-cyclic phosphates 5' to the cleaved bond (By similarity).

See Also

3vcb, resolution 2.40Å

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