Viral RNA annealing activities of the nucleocapsid protein of Moloney murine leukemia virus are zinc independent.
The zinc fingers of retroviral gag nucleocapsid proteins (NC) are required for the specific packaging of the dimeric RNA genome into virions. In vitro, NC proteins activate both dimerization of viral RNA and annealing of the replication primer tRNA onto viral RNA, two reactions necessary for the production of infectious virions. In this study the role of the zinc finger of Moloney murine leukemia virus (MoMuLV) NCp10 in RNA binding and annealing activities was investigated through modification or replacement of residues involved in zinc coordination. These alterations did not affect the ability of NCp10 to bind RNA and promote RNA annealing in vitro, despite a complete loss of zinc affinity. However mutation of two conserved lysine residues adjacent to the finger motif reduced both RNA binding and annealing activities of NCp10. These findings suggest that the complexed NC zinc finger is not directly involved in RNA-protein interactions but more probably in a zinc dependent conformation of NC protein modulating viral protein-protein interactions, essential to the process of viral RNA selection and virion assembly. Then the NC zinc finger may cooperate to select the viral RNA genome to be packaged into virions.
Prats
A C
AC
Centre de Recherche de Biochimie et Génétique Cellulaires du CNRS, Toulouse, France.
Housset
V
V
de Billy
G
G
Cornille
F
F
Prats
H
H
Roques
B
B
Darlix
J L
JL
D000478:Alkylation; D000595:Amino Acid Sequence; D001483:Base Sequence; D002213:Capsid / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D003545:Cysteine / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D007700:Kinetics; D008239:Lysine / Q000378:metabolism; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000235:genetics / Q000502:physiology*; D009154:Mutation / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D012361:RNA, Transfer, Pro / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism*; D014758:Viral Core Proteins / Q000235:genetics / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism; D016335:Zinc Fingers / Q000502:physiology*
D012361:RNA, Transfer, Pro; D012367:RNA, Viral; D014758:Viral Core Proteins; D015032:Zinc; D008239:Lysine; D003545:Cysteine