7ck5

Solution structure of 28 amino acid polypeptide (354-381) in Plantago asiatica mosaic virus replicase bound to SDS micelle

spinqualitylabelsall models 7ck5 Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 7ck5 is a 1 chain structure with sequence from Plantago asiatica mosaic virus. Full experimental information is available from OCA. For a guided tour on the structure components use FirstGlance. Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function B1B578_P1AMV RNA replication. The central part of this protein possibly functions as an ATP-binding helicase.[ARBA:ARBA00003129] Publication Abstract from PubMed Characterized positive-strand RNA viruses replicate in association with intracellular membranes. Regarding viruses in the genus Potexvirus, the mechanism by which their RNA-dependent RNA polymerase (replicase) associates with membranes is understudied. Here, by membrane flotation analyses of the replicase of Plantago asiatica mosaic potexvirus (PlAMV), we identified a region in the methyltransferase (MET) domain as a membrane association determinant. An amphipathic alpha-helix was predicted downstream from the core region of the MET domain, and hydrophobic amino acid residues were conserved in the helical sequences in replicases of other potexviruses. Nuclear magnetic resonance (NMR) analysis confirmed the amphipathic alpha-helical configuration and unveiled a kink caused by a highly conserved proline residue in the alpha-helix. Substitution of this proline residue and other hydrophobic and charged residues in the amphipathic alpha-helix abolished PlAMV replication. Ectopic expression of a green fluorescent protein (GFP) fusion with the entire MET domain resulted in the formation of a large perinuclear complex, where virus replicase and RNA colocated during virus infection. Except for the proline substitution, the amino acid substitutions in the alpha-helix that abolished virus replication also prevented the formation of the large perinuclear complex by the respective GFP-MET fusion. Small intracellular punctate structures were observed for all GFP-MET fusions, and in vitro high-molecular-weight complexes were formed by both replication-competent and -incompetent viral replicons and thus were not sufficient for replication competence. We discuss the roles of the potexvirus-specific, proline-kinked amphipathic helical structure in virus replication and intracellular large complex and punctate structure formation. IMPORTANCE RNA viruses characteristically associate with intracellular membranes during replication. Although virus replicases are assumed to possess membrane-targeting properties, their membrane association domains generally remain unidentified or poorly characterized. Here, we identified a proline-kinked amphipathic alpha-helix structure downstream from the methyltransferase core domain of PlAMV replicase as a membrane association determinant. This helical sequence, which includes the proline residue, was conserved among potexviruses and related viruses in the order Tymovirales. Substitution of the proline residue, but not the other residues necessary for replication, allowed formation of a large perinuclear complex within cells resembling those formed by PlAMV replicase and RNA during virus replication. Our results demonstrate the role of the amphipathic alpha-helix in PlAMV replicase in a perinuclear complex formation and virus replication and that perinuclear complex formation by the replicase alone will not necessarily indicate successful virus replication. Identification of a Proline-Kinked Amphipathic alpha-Helix Downstream from the Methyltransferase Domain of a Potexvirus Replicase and Its Role in Virus Replication and Perinuclear Complex Formation.,Komatsu K, Sasaki N, Yoshida T, Suzuki K, Masujima Y, Hashimoto M, Watanabe S, Tochio N, Kigawa T, Yamaji Y, Oshima K, Namba S, Nelson RS, Arie T J Virol. 2021 Sep 27;95(20):e0190620. doi: 10.1128/JVI.01906-20. Epub 2021 Aug 4. PMID:34346768 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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