Conserved cysteine and histidine residues of the avian myeloblastosis virus nucleocapsid protein are essential for viral replication but are not "zinc-binding fingers".
The nucleocapsid protein from the Rous sarcoma virus has two regions of sequence with the motif Cys-Xaa-Xaa-Cys-Xaa-Xaa-Xaa-Gly-His-Xaa-Xaa-Xaa-Cys. All retrovirus nucleocapsid proteins contain one or two of these motifs, and they represent the only conserved sequences among these proteins. Sequence analysis of nucleocapsid from avian myeloblastosis virus shows that it also contains two Cys-His sequences and, in fact, differs from the Rous sarcoma nucleocapsid protein only in three residues near the carboxyl terminus. The hypothesized role of the conserved cysteines and histidines as zinc ligands was tested experimentally. No tightly bound metal ions were detected for avian myeloblastosis nucleocapsid protein, and the molar amount of zinc in virions was less by a factor of 50 than that of the nucleocapsid protein. Added Zn2+ did not significantly affect nucleocapsid binding to poly(ethenoadenylic acid) or its secondary structure, as determined from circular dichroism. Nevertheless, the conserved cysteine and histidine residues of the Rous sarcoma (Prague-C strain) nucleocapsid protein are essential for fully functional virus, as shown by the fact that single-site substitutions of five of the six conserved cysteines and either of the two histidine residues blocked viral replication.
Jentoft
J E
JE
Department of Biochemistry, School of Medicine, Case Western Reserve University, Cleveland, OH 44106.
Smith
L M
LM
Fu
X D
XD
Johnson
M
M
Leis
J
J
D001354:Avian Leukosis Virus / Q000032:analysis*; D009189:Avian Myeloblastosis Virus / Q000032:analysis*; D001483:Base Sequence; D002213:Capsid / Q000032:analysis*; D002942:Circular Dichroism; D003545:Cysteine / Q000032:analysis*; D006639:Histidine / Q000032:analysis*; D009154:Mutation; D011487:Protein Conformation; D014758:Viral Core Proteins / Q000032:analysis*; D014779:Virus Replication; D015032:Zinc / Q000378:metabolism*
D014758:Viral Core Proteins; D006639:Histidine; D015032:Zinc; D003545:Cysteine