Phosphorylation of Marburg virus VP30 at serines 40 and 42 is critical for its interaction with NP inclusions.
The Marburg virus (MBGV) nucleocapsid complex is composed of four viral proteins (NP, L, VP35, and VP30) and the negative-strand nonsegmented genomic RNA. NP, L, and VP35 are functionally conserved among the order Mononegavirales, whereas VP30, a phosphoprotein, represents a filovirus-specific nucleocapsid protein. In the present paper, we have characterized the localization and function of VP30 phosphorylation. The main phosphorylation sites are represented by seven serine residues in the region of amino acid 40 to 51 of VP30. Additionally, trace amounts of phosphothreonine were detected. Substitution of serine residues 40 and 42 by alanine abolished the interaction of VP30 with NP-induced inclusion bodies, which contain nucleocapsid-like structures formed by NP. Substitution of the other phosphoserine residues had little effect on this interaction. Replacement of the introduced alanine residues 40 and 42 by aspartate restored the interaction between VP30 and the NP inclusions pointing to the importance of negative charges at these particular positions.
Modrof
J
J
Institut für Virologie der Philipps-Universität Marburg, Robert-Koch-Str. 17, 35037 Marburg, Germany.
Möritz
C
C
Kolesnikova
L
L
Konakova
T
T
Hartlieb
B
B
Randolf
A
A
Mühlberger
E
E
Becker
S
S
D000469:Alkaline Phosphatase / Q000378:metabolism; D019943:Amino Acid Substitution; D000818:Animals; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D005561:Formates / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D007181:Inclusion Bodies, Viral / Q000378:metabolism*; D016253:Microscopy, Immunoelectron; D016297:Mutagenesis, Site-Directed; D019590:Nucleocapsid Proteins / Q000378:metabolism*; D019606:Phosphoamino Acids / Q000378:metabolism; D010766:Phosphorylation; D012694:Serine / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014162:Transfection; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D005561:Formates; D019590:Nucleocapsid Proteins; D019606:Phosphoamino Acids; C094001:VP30 protein, Marburg virus; D014764:Viral Proteins; C030544:formic acid; D012694:Serine; D000469:Alkaline Phosphatase