title: Effect of dimerizing domains and basic residues on in vitro and in vivo assembly of Mason-Pfizer monkey virus and human immunodeficiency virus.
abstract: Assembly of immature retroviral particles is a complex process involving interactions of several specific domains of the Gag polyprotein localized mainly within capsid protein (CA), spacer peptide (SP), and nucleocapsid protein (NC). In the present work we focus on the contribution of NC to the oligomerization of CA leading to assembly of Mason-Pfizer monkey virus (M-PMV) and HIV-1. Analyzing in vitro assembly of substitution and deletion mutants of DeltaProCANC, we identified a "spacer-like" sequence (NC(15)) at the M-PMV NC N terminus. This NC(15) domain is indispensable for the assembly and cannot be replaced with oligomerization domains of GCN4 or CREB proteins. Although the M-PMV NC(15) occupies a position analogous to that of the HIV-1 spacer peptide, it could not be replaced by the latter one. To induce the assembly, both M-PMV NC(15) and HIV-1 SP1 must be followed by a short peptide that is rich in basic residues. This region either can be specific, i.e., derived from the downstream NC sequence, or can be a nonspecific positively charged peptide. However, it cannot be replaced by heterologous interaction domains either from GCN4 or from CREB. In summary, we report here a novel M-PMV spacer-like domain that is functionally similar to other retroviral spacer peptides and contributes to the assembly of immature-virus-like particles.
authors.affiliation: Institute of Organic Chemistry and Biochemistry IOCB Research Centre & Gilead Sciences, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 10, Prague, Czech Republic.
authors.forename: Karolína
authors.initials: K
authors.lastname: Bohmová
authors.forename: Romana
authors.initials: R
authors.lastname: Hadravová
authors.forename: Jitka
authors.initials: J
authors.lastname: Stokrová
authors.forename: Roman
authors.initials: R
authors.lastname: Tuma
authors.forename: Tomás
authors.initials: T
authors.lastname: Ruml
authors.forename: Iva
authors.initials: I
authors.lastname: Pichová
authors.forename: Michaela
authors.initials: M
authors.lastname: Rumlová
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000648:ultrastructure / Q000821:virology; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D016093:Mason-Pfizer monkey virus / Q000235:genetics* / Q000502:physiology* / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission; D008969:Molecular Sequence Data; D016296:Mutagenesis; D019590:Nucleocapsid Proteins / Q000737:chemistry* / Q000235:genetics* / Q000502:physiology; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D019065:Virus Assembly / Q000235:genetics / Q000502:physiology
chemical_list: D017931:DNA Primers; D004279:DNA, Viral; D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins