title: Stabilization of the β-hairpin in Mason-Pfizer monkey virus capsid protein- a critical step for infectivity.
abstract: BACKGROUND
  Formation of a mature core is a crucial event for infectivity of retroviruses such as Mason-Pfizer monkey virus (M-PMV). The process is triggered by proteolytic cleavage of the polyprotein precursor Gag, which releases matrix, capsid (CA), and nucleocapsid proteins. Once released, CA assembles to form a mature core - a hexameric lattice protein shell that protects retroviral genomic RNA. Subtle conformational changes within CA induce the transition from the immature lattice to the mature lattice. Upon release from the precursor, the initially unstructured N-terminus of CA is refolded to form a β-hairpin stabilized by a salt bridge between the N-terminal proline and conserved aspartate. Although the crucial role of the β-hairpin in the mature core assembly has been confirmed, its precise structural function remains poorly understood.
  
  
  RESULTS
  Based on a previous NMR analysis of the N-terminal part of M-PMV CA, which suggested the role of additional interactions besides the proline-aspartate salt bridge in stabilization of the β-hairpin, we introduced a series of mutations into the CA sequence. The effect of the mutations on virus assembly and infectivity was analyzed. In addition, the structural consequences of selected mutations were determined by NMR spectroscopy. We identified a network of interactions critical for proper formation of the M-PMV core. This network involves residue R14, located in the N-terminal β-hairpin; residue W52 in the loop connecting helices 2 and 3; and residues Q113, Q115, and Y116 in helix 5.
  
  
  CONCLUSIONS
  Combining functional and structural analyses, we identified a network of supportive interactions that stabilize the β-hairpin in mature M-PMV CA.
authors.forename: Martin
authors.initials: M
authors.lastname: Obr
authors.forename: Romana
authors.initials: R
authors.lastname: Hadravová
authors.forename: Michal
authors.initials: M
authors.lastname: DoleŽal
authors.forename: Ivana
authors.initials: I
authors.lastname: KříŽová
authors.forename: Veronika
authors.initials: V
authors.lastname: Papoušková
authors.forename: Lukáš
authors.initials: L
authors.lastname: Zídek
authors.forename: Richard
authors.initials: R
authors.lastname: Hrabal
authors.forename: Tomáš
authors.initials: T
authors.lastname: Ruml
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, v,v,i,, IOCB & Gilead Research Center, Flemingovo nám, 2, Prague, 166 10, Czech Republic. rumlova@uochb.cas.cz.
authors.forename: Michaela
authors.initials: M
authors.lastname: Rumlová
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D036022:Capsid Proteins / Q000378:metabolism*; D002460:Cell Line; D057809:HEK293 Cells; D006801:Humans; D016093:Mason-Pfizer monkey virus / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D017433:Protein Structure, Secondary / Q000235:genetics*; D016097:Simian Acquired Immunodeficiency Syndrome / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000235:genetics / Q000378:metabolism; D019065:Virus Assembly / Q000235:genetics
chemical_list: D036022:Capsid Proteins