Genome polyprotein Recombinant Protein | HCV4_gp1 recombinant protein

Recombinant Hepatitis C virus genotype 4a Genome polyprotein

Cat. Num. AAA1079380

• Purity: Greater or equal to 85% purity as determined by SDS-PAGE.
• Synonyms: Genome polyprotein; Recombinant Hepatitis C virus genotype 4a Genome polyprotein; Recombinant Genome polyprotein; Genome polyprotein Cleaved into the following 11 chains: 1. Core protein p21; Capsid protein C p21 Core protein p19 Envelope glycoprotein E1; gp32 gp35 Envelope glycoprotein E2; NS1 gp68 gp70 p7 Protease NS2-3; p23 EC= 3.4.; HCV4_gp1 recombinant protein

• Host: E Coli or Yeast or Baculovirus or Mammalian Cell
• Purity/Purification: Greater or equal to 85% purity as determined by SDS-PAGE.
• Form/Format: Lyophilized or liquid (Format to be determined during the manufacturing process)
• Sequence Positions: 190-358aa; Partial
• Sequence: SAVNYRNVSGIYHVTNDCPNSSIVYEADHHIMHLPGCVPCVREGNQSRCWVALTPTVAAPYIGAPLESLRSHVDLMVGAATVCSGLYIGDLCGGLFLVGQMFSFRPRRHWTTQDCNCSIYTGHITGHRMAWDMMMNWSPTTTLVLAQVMRIPTTLVDLLSGGHWGVLVG

• Species: Hepatitis C virus genotype 4a (isolate ED43) (HCV)
• Preparation and Storage: Store at -20 degree C. For extended storage, store at -20 or -80 degree C.

NCBI and Uniprot Product Information

• NCBI GI #: 157781209
• NCBI GeneID: 11027168
• NCBI Accession #: YP_001469632.1
• NCBI GenBank Nucleotide #: NC_009825.1
• UniProt Accession #: O39929
• Molecular Weight: 327,599 Da
• NCBI Official Full Name: HCV polyprotein
• NCBI Official Symbol: HCV4_gp1
• NCBI Protein Information: HCV polyprotein
• UniProt Protein Name: Genome polyprotein
• Protein Family: Genome polyprotein
• UniProt Gene Name: p23
• UniProt Synonym Gene Names: NS4A; NS4B; NS5A
• UniProt Entry Name: POLG_HCVED

Uniprot Description

Function: Core protein packages viral RNA to form a viral nucleocapsid, and promotes virion budding. Modulates viral translation initiation by interacting with HCV IRES and 40S ribosomal subunit. Also regulates many host cellular functions such as signaling pathways and apoptosis. Prevents the establishment of cellular antiviral state by blocking the interferon-alpha/beta (IFN-alpha/beta) and IFN-gamma signaling pathways and by inducing human STAT1 degradation. Thought to play a role in virus-mediated cell transformation leading to hepatocellular carcinomas. Interacts with, and activates STAT3 leading to cellular transformation. May repress the promoter of p53, and sequester CREB3 and SP110 isoform 3/Sp110b in the cytoplasm. Also represses cell cycle negative regulating factor CDKN1A, thereby interrupting an important check point of normal cell cycle regulation. Targets transcription factors involved in the regulation of inflammatory responses and in the immune response: suppresses NK-kappaB activation, and activates AP-1. Could mediate apoptotic pathways through association with TNF-type receptors TNFRSF1A and LTBR, although its effect on death receptor-induced apoptosis remains controversial. Enhances TRAIL mediated apoptosis, suggesting that it might play a role in immune-mediated liver cell injury. Seric core protein is able to bind C1QR1 at the T-cell surface, resulting in down-regulation of T-lymphocytes proliferation. May transactivate human MYC, Rous sarcoma virus LTR, and SV40 promoters. May suppress the human FOS and HIV-1 LTR activity. Alters lipid metabolism by interacting with hepatocellular proteins involved in lipid accumulation and storage. Core protein induces up-regulation of FAS promoter activity, and thereby probably contributes to the increased triglyceride accumulation in hepatocytes (steatosis)

By similarity.E1 and E2 glycoproteins form a heterodimer that is involved in virus attachment to the host cell, virion internalization through clathrin-dependent endocytosis and fusion with host membrane. E1/E2 heterodimer binds to human LDLR, CD81 and SCARB1/SR-BI receptors, but this binding is not sufficient for infection, some additional liver specific cofactors may be needed. The fusion function may possibly be carried by E1. E2 inhibits human EIF2AK2/PKR activation, preventing the establishment of an antiviral state. E2 is a viral ligand for CD209/DC-SIGN and CLEC4M/DC-SIGNR, which are respectively found on dendritic cells (DCs), and on liver sinusoidal endothelial cells and macrophage-like cells of lymph node sinuses. These interactions allow capture of circulating HCV particles by these cells and subsequent transmission to permissive cells. DCs act as sentinels in various tissues where they entrap pathogens and convey them to local lymphoid tissue or lymph node for establishment of immunity. Capture of circulating HCV particles by these SIGN+ cells may facilitate virus infection of proximal hepatocytes and lymphocyte subpopulations and may be essential for the establishment of persistent infection

By similarity.P7 seems to be a heptameric ion channel protein (viroporin) and is inhibited by the antiviral drug amantadine. Also inhibited by long-alkyl-chain iminosugar derivatives. Essential for infectivity

By similarity.Protease NS2-3 is a cysteine protease responsible for the autocatalytic cleavage of NS2-NS3. Seems to undergo self-inactivation following maturation

By similarity.NS3 displays three enzymatic activities: serine protease, NTPase and RNA helicase. NS3 serine protease, in association with NS4A, is responsible for the cleavages of NS3-NS4A, NS4A-NS4B, NS4B-NS5A and NS5A-NS5B. NS3/NS4A complex also prevents phosphorylation of human IRF3, thus preventing the establishment of dsRNA induced antiviral state. NS3 RNA helicase binds to RNA and unwinds dsRNA in the 3' to 5' direction, and likely RNA stable secondary structure in the template strand. Cleaves and inhibits the host antiviral protein MAVS

By similarity.NS4B induces a specific membrane alteration that serves as a scaffold for the virus replication complex. This membrane alteration gives rise to the so-called ER-derived membranous web that contains the replication complex

By similarity.NS5A is a component of the replication complex involved in RNA-binding. Its interaction with Human VAPB may target the viral replication complex to vesicles. Down-regulates viral IRES translation initiation. Mediates interferon resistance, presumably by interacting with and inhibiting human EIF2AK2/PKR. Seems to inhibit apoptosis by interacting with BIN1 and FKBP8. The hyperphosphorylated form of NS5A is an inhibitor of viral replication

By similarity.NS5B is a RNA-dependent RNA polymerase that plays an essential role in the virus replication

By similarity.

Catalytic activity: Hydrolysis of four peptide bonds in the viral precursor polyprotein, commonly with Asp or Glu in the P6 position, Cys or Thr in P1 and Ser or Ala in P1'.Nucleoside triphosphate + RNA(n) = diphosphate + RNA(n+1).NTP + H2O = NDP + phosphate.ATP + H2O = ADP + phosphate.

Cofactor: Binds 1 zinc ion per NS3 protease domain

By similarity.Binds 1 zinc ion per NS5A N-terminal domain

By similarity.

Enzyme regulation: Activity of auto-protease NS2-3 is dependent on zinc ions and completely inhibited by EDTA. Serine protease NS3 is also activated by zinc ions

By similarity.

Subunit structure: Core protein is a homomultimer that binds the C-terminal part of E1 and interacts with numerous cellular proteins. Interaction with human STAT1 SH2 domain seems to result in decreased STAT1 phosphorylation, leading to decreased IFN-stimulated gene transcription. In addition to blocking the formation of phosphorylated STAT1, the core protein also promotes ubiquitin-mediated proteasome-dependent degradation of STAT1. Interacts with, and constitutively activates human STAT3. Associates with human LTBR and TNFRSF1A receptors and possibly induces apoptosis. Binds to human SP110 isoform 3/Sp110b, HNRPK, C1QR1, YWHAE, UBE3A/E6AP, DDX3X, APOA2 and RXRA proteins. Interacts with human CREB3 nuclear transcription protein, triggering cell transformation. May interact with human p53. Also binds human cytokeratins KRT8, KRT18, KRT19 and VIM (vimentin). E1 and E2 glycoproteins form a heterodimer that binds to human LDLR, CLDN1, CD81 and SCARB1 receptors. E2 binds and inhibits human EIF2AK2/PKR. Also binds human CD209/DC-SIGN and CLEC4M/DC-SIGNR. p7 forms a homoheptamer in vitro. NS2 forms a homodimer containing a pair of composite active sites at the dimerization interface. NS2 seems to interact with all other non-structural (NS) proteins. NS4A interacts with NS3 serine protease and stabilizes its folding. NS3-NS4A complex is essential for the activation of the latter and allows membrane anchorage of NS3. NS3 interacts with human TANK-binding kinase TBK1 and MAVS. NS4B and NS5A form homodimers and seem to interact with all other non-structural (NS) proteins. NS5A also interacts with human EIF2AK2/PKR, FKBP8, GRB2, BIN1, PIK3R1, SRCAP, VAPB and with most Src-family kinases. NS5B is a homooligomer and interacts with human VAPB, HNRNPA1 and SEPT6

By similarity. Ref.4

Subcellular location: Core protein p21: Host endoplasmic reticulum membrane; Single-pass membrane protein

By similarity. Host mitochondrion membrane; Single-pass type I membrane protein

By similarity. Host lipid droplet

By similarity. Note: The C-terminal transmembrane domain of core protein p21 contains an ER signal leading the nascent polyprotein to the ER membrane. Only a minor proportion of core protein is present in the nucleus and an unknown proportion is secreted. Ref.3Core protein p19: Virion

By similarity. Host cytoplasm

By similarity. Host nucleus

By similarity. Secreted

By similarity Ref.3. Envelope glycoprotein E1: Virion membrane; Single-pass type I membrane protein

Potential. Host endoplasmic reticulum membrane; Single-pass type I membrane protein

By similarity. Note: The C-terminal transmembrane domain acts as a signal sequence and forms a hairpin structure before cleavage by host signal peptidase. After cleavage, the membrane sequence is retained at the C-terminus of the protein, serving as ER membrane anchor. A reorientation of the second hydrophobic stretch occurs after cleavage producing a single reoriented transmembrane domain. These events explain the final topology of the protein. ER retention of E1 is leaky and, in overexpression conditions, only a small fraction reaches the plasma membrane. Ref.3Envelope glycoprotein E2: Virion membrane; Single-pass type I membrane protein

Potential. Host endoplasmic reticulum membrane; Single-pass type I membrane protein

By similarity. Note: The C-terminal transmembrane domain acts as a signal sequence and forms a hairpin structure before cleavage by host signal peptidase. After cleavage, the membrane sequence is retained at the C-terminus of the protein, serving as ER membrane anchor. A reorientation of the second hydrophobic stretch occurs after cleavage producing a single reoriented transmembrane domain. These events explain the final topology of the protein. ER retention of E2 is leaky and, in overexpression conditions, only a small fraction reaches the plasma membrane. Ref.3p7: Host endoplasmic reticulum membrane; Multi-pass membrane protein

By similarity Ref.3. Host cell membrane

By similarity. Note: The C-terminus of p7 membrane domain acts as a signal sequence. After cleavage by host signal peptidase, the membrane sequence is retained at the C-terminus of the protein, serving as ER membrane anchor. Only a fraction localizes to the plasma membrane. Ref.3Protease NS2-3: Host endoplasmic reticulum membrane; Multi-pass membrane protein

Potential Ref.3. Serine protease NS3: Host endoplasmic reticulum membrane; Peripheral membrane protein

By similarity. Note: NS3 is associated to the ER membrane through its binding to NS4A. Ref.3Non-structural protein 4A: Host endoplasmic reticulum membrane; Single-pass type I membrane protein

Potential. Note: Host membrane insertion occurs after processing by the NS3 protease. Ref.3Non-structural protein 4B: Host endoplasmic reticulum membrane; Multi-pass membrane protein

By similarity Ref.3. Non-structural protein 5A: Host endoplasmic reticulum membrane; Peripheral membrane protein

By similarity. Host cytoplasm › host perinuclear region

By similarity. Host mitochondrion

By similarity. Note: Host membrane insertion occurs after processing by the NS3 protease. Ref.3RNA-directed RNA polymerase: Host endoplasmic reticulum membrane; Single-pass type I membrane protein

Potential. Note: Host membrane insertion occurs after processing by the NS3 protease. Ref.3

Domain: The transmembrane regions of envelope E1 and E2 glycoproteins are involved in heterodimer formation, ER localization, and assembly of these proteins. Envelope E2 glycoprotein contain a highly variable region called hypervariable region 1 (HVR1). E2 also contains two segments involved in CD81-binding. HVR1 is implicated in the SCARB1-mediated cell entry. CD81-binding regions may be involved in sensitivity and/or resistance to IFN-alpha therapy

By similarity.The N-terminus of NS5A act as membrane anchor. The central part of NS5A seems to be intrinsically disordered and interacts with NS5B and host PKR

By similarity.The SH3-binding domain of NS5A is involved in the interaction with human Bin1, GRB2 and Src-family kinases

By similarity.The N-terminal one-third of serine protease NS3 contains the protease activity. This region contains a zinc atom that does not belong to the active site, but may play a structural rather than a catalytic role. This region is essential for the activity of protease NS2-3, maybe by contributing to the folding of the latter. The helicase activity is located in the C-terminus of NS3

By similarity.

Post-translational modification: Specific enzymatic cleavages in vivo yield mature proteins. The structural proteins, core, E1, E2 and p7 are produced by proteolytic processing by host signal peptidases. The core protein is synthesized as a 21 kDa precursor which is retained in the ER membrane through the hydrophobic signal peptide. Cleavage by the signal peptidase releases the 19 kDa mature core protein. The other proteins (p7, NS2-3, NS3, NS4A, NS4B, NS5A and NS5B) are cleaved by the viral proteases

By similarity.Envelope E1 and E2 glycoproteins are highly N-glycosylated

By similarity.Core protein is phosphorylated by host PKC and PKA

By similarity.NS5A is phosphorylated in a basal form termed p56. p58 is an hyperphosphorylated form of p56. p56 and p58 coexist in the cell in roughly equivalent amounts. Hyperphosphorylation is dependent on the presence of NS4A. Human AKT1, RPS6KB1/p70S6K, MAP2K1/MEK1, MAP2K6/MKK6 and CSNK1A1/CKI-alpha kinases may be responsible for NS5A phosphorylation

By similarity.NS4B is palmitoylated. This modification may play a role in its polymerization or in protein-protein interactions

By similarity.The N-terminus of a fraction of NS4B molecules seems to be relocated post-translationally from the cytoplasm to the ER lumen, with a 5th transmembrane segment. The C-terminus of NS2 may be lumenal with a fourth transmembrane segment

By similarity.Core protein is ubiquitinated; mediated by UBE3A and leading to core protein subsequent proteasomal degradation

By similarity.

Miscellaneous: Cell culture adaptation of the virus leads to mutations in NS5A, reducing its inhibitory effect on replication

By similarity.Core protein exerts viral interference on hepatitis B virus when HCV and HBV coinfect the same cell, by suppressing HBV gene expression, RNA encapsidation and budding

By similarity.

Sequence similarities: Belongs to the hepacivirus polyprotein family.Contains 1 helicase ATP-binding domain.Contains 1 peptidase C18 domain.Contains 1 peptidase S29 domain.Contains 1 RdRp catalytic domain.

Caution: The core gene probably also codes for alternative reading frame proteins (ARFPs). Many functions depicted for the core protein might belong to the ARFPs.