Zinc-binding properties of Junín virus nucleocapsid protein.
The arenavirus nucleocapsid protein (N) is a highly basic 63 kDa protein with a dual function during the virus life-cycle. First, it is involved in essential steps of genome replication, promoting the synthesis of the full-length antigenomic copy of S RNA, and second it associates with the genomic RNA to form the nucleocapsid. We have expressed the N protein of Junín virus in E. coli and shown that it binds zinc in vitro. This property is in agreement with the presence in the carboxy-terminal region of the N protein of the CX(2)HX(23)CX(4)C sequence, which resembles a classical zinc-finger motif. The specificity for zinc binding was demonstrated by competition with other divalent metal ions. The ability of the predicted motif to bind zinc was established by analysis of a series of N mutants, including truncated variants and amino acid substitutions. In addition, alternative zinc-binding sites were found.
Tortorici
M Alejandra
MA
Instituto de Bioquímica y Biología Molecular, Depto de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, Calles 47 y 115, 1900 La Plata, Buenos Aires, Argentina1.
Ghiringhelli
P Daniel
PD
Laboratorio de Ingeniería Genética y Biología Celular y Molecular, Departamento de Ciencia y Tecnología, Centro de Estudios e Investigaciones, Universidad Nacional de Quilmes, Roque Saenz Peña 180, 1876 Bernal, Buenos Aires, Argentina2.
Lozano
Mario E
ME
Laboratorio de Ingeniería Genética y Biología Celular y Molecular, Departamento de Ciencia y Tecnología, Centro de Estudios e Investigaciones, Universidad Nacional de Quilmes, Roque Saenz Peña 180, 1876 Bernal, Buenos Aires, Argentina2.
Albariño
César G
CG
Instituto de Bioquímica y Biología Molecular, Depto de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, Calles 47 y 115, 1900 La Plata, Buenos Aires, Argentina1.
Romanowski
Víctor
V
Laboratorio de Ingeniería Genética y Biología Celular y Molecular, Departamento de Ciencia y Tecnología, Centro de Estudios e Investigaciones, Universidad Nacional de Quilmes, Roque Saenz Peña 180, 1876 Bernal, Buenos Aires, Argentina2.|Instituto de Bioquímica y Biología Molecular, Depto de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, Calles 47 y 115, 1900 La Plata, Buenos Aires, Argentina1.
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D004926:Escherichia coli / Q000235:genetics; D014644:Genetic Variation; D005822:Genetic Vectors; D018051:Junin virus / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D019590:Nucleocapsid Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis; D016415:Sequence Alignment; D015032:Zinc / Q000378:metabolism*
D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins; D015032:Zinc