Asymmetric packaging of polymerases within vesicular stomatitis virus.
Vesicular stomatitis virus (VSV) is a prototypic negative sense single-stranded RNA virus. The bullet-shape appearance of the virion results from tightly wound helical turns of the nucleoprotein encapsidated RNA template (N-RNA) around a central cavity. Transcription and replication require polymerase complexes, which include a catalytic subunit L and a template-binding subunit P. L and P are inferred to be in the cavity, however lacking direct observation, their exact position has remained unclear. Using super-resolution fluorescence imaging and atomic force microscopy (AFM) on single VSV virions, we show that L and P are packaged asymmetrically towards the blunt end of the virus. The number of L and P proteins varies between individual virions and they occupy 57 ± 12 nm of the 150 nm central cavity of the virus. Our finding positions the polymerases at the opposite end of the genome with respect to the only transcriptional promoter.
Dept. of Physics and Astronomy, University of Utah, United States; Center for Cell and Genome Science, University of Utah, United States.
Jeffery
J
Hodges
Xiaolin
X
Tang
Michael B
MB
Landesman
John B
JB
Ruedas
Anil
A
Ghimire
Manasa V
MV
Gudheti
Jacques
J
Perrault
Erik M
EM
Jorgensen
Jordan M
JM
Gerton
Saveez
S
Saffarian
D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D018625:Microscopy, Atomic Force; D008856:Microscopy, Fluorescence; D012367:RNA, Viral / Q000235:genetics; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics* / Q000648:ultrastructure; D014764:Viral Proteins / Q000378:metabolism*; D019065:Virus Assembly / Q000502:physiology*
D012367:RNA, Viral; D014764:Viral Proteins; D012321:DNA-Directed RNA Polymerases; C512399:L polymerase protein, Vesicular stomatitis-Indiana virus
AFM; L protein; P protein; PALM; Polymerase; RdRP; STORM; VSV; Viral replication