NMR structure of the N-terminal coiled coil domain of the Andes hantavirus nucleocapsid protein.
The hantaviruses are emerging infectious viruses that in humans can cause a cardiopulmonary syndrome or a hemorrhagic fever with renal syndrome. The nucleocapsid (N) is the most abundant viral protein, and during viral assembly, the N protein forms trimers and packages the viral RNA genome. Here, we report the NMR structure of the N-terminal domain (residues 1-74, called N1-74) of the Andes hantavirus N protein. N1-74 forms two long helices (alpha1 and alpha2) that intertwine into a coiled coil domain. The conserved hydrophobic residues at the helix alpha1-alpha2 interface stabilize the coiled coil; however, there are many conserved surface residues whose function is not known. Site-directed mutagenesis, CD spectroscopy, and immunocytochemistry reveal that a point mutation in the conserved basic surface formed by Arg22 or Lys26 lead to antibody recognition based on the subcellular localization of the N protein. Thus, Arg22 and Lys26 are likely involved in a conformational change or molecular recognition when the N protein is trafficked from the cytoplasm to the Golgi, the site of viral assembly and maturation.
Wang
Yu
Y
Department of Molecular Biosciences, University of Kansas, Lawrence, Kansas 66045, USA.
Boudreaux
Daniel M
DM
Estrada
D Fernando
DF
Egan
Chet W
CW
St Jeor
Stephen C
SC
De Guzman
Roberto N
RN
D000818:Animals; D001120:Arginine / Q000737:chemistry; D019556:COS Cells; D002522:Chlorocebus aethiops; D002942:Circular Dichroism; D003593:Cytoplasm / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D006476:Orthohantavirus / Q000378:metabolism*; D008239:Lysine / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D016297:Mutagenesis, Site-Directed; D019590:Nucleocapsid Proteins / Q000737:chemistry*; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary
D019590:Nucleocapsid Proteins; D001120:Arginine; D008239:Lysine