Mutational analysis of positively charged amino acid residues of Uukuniemi phlebovirus nucleocapsid protein.
The aim of this study was to evaluate the contribution of positively charged amino acid residues for the Uukuniemi virus (UUKV) N protein functionality. Based on phlebovirus nucleocapsid (N) protein alignments and 3D-structure predictions of UUKV N protein, 14 positively charged residues were chosen as targets for the mutagenesis. The impact of mutations to the N protein functionality was analyzed using minigenome-, virus-like particle-, and mammalian two-hybrid-assays. Seven of the mutations affected the functional competence in all three assays, while others had milder impact or no impact at all. In the 3D-model of UUKV N protein, five of the affected residues, R61, R64, R73, R98 and R115, were located either within or in close proximity to the central cavity that could potentially bind RNA.
Research Programs Unit, Infection Biology Research Program, Department of Virology, Haartman Institute, PO Box 21, University of Helsinki, 00014 Helsinki, Finland. anna.katz@helsinki.fi
Anna
A
Katz
Alexander N
AN
Freiberg
Vera
V
Backström
Liisa
L
Holm
Antti
A
Vaheri
Ramon
R
Flick
Alexander
A
Plyusnin
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004252:DNA Mutational Analysis; D008969:Molecular Sequence Data; D009154:Mutation; D019590:Nucleocapsid Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016856:Phlebovirus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D018066:Uukuniemi virus / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D000596:Amino Acids; D019590:Nucleocapsid Proteins