Composition and mass of the bacteriophage phi29 prohead and virion.
The protein composition of the Bacillus subtilis bacteriophage phi29 prohead and virion was determined by combustion of gel bands of (3)H-labeled proteins. Copy numbers of individual proteins were calculated relative to the 12 copies of the head-tail connector protein. The mean numbers of copies of the major capsid protein in the prohead and virion were 241 and 218, respectively, approaching the 235 copies determined previously by cryoelectron microscopy. The mean numbers of copies of the dimeric head fiber on the prohead and virion were 24 and 31, respectively, demonstrating partial occupancy of the 55 fiber binding sites. Measured copies of neck and tail proteins in the virion included 11 of the lower collar, 58 of the appendage, and 9 of the tail; if the true copies of these proteins are 12, 60, and 9, respectively, the entire neck and tail of phi29 has quasi-sixfold symmetry. The mass of the fiberless prohead with pRNA was about 14.2 MDa, and the mass of the prohead determined by scanning transmission electron microscopy was consistent with the biochemical data. The mass of the fiberless virion containing the 12.8-MDa DNA genome was about 30.4 MDa. A full complement of dimeric fibers on the prohead or virion would increase the mass of the particle by about 3.2 MDa. The data complement studies relating the structure of phi29 components to dynamic functions in morphogenesis and infection.
Peterson
C
C
Department of Oral Science, University of Minnesota, Minneapolis, Minnesota 55455, USA.
Simon
M
M
Hodges
J
J
Mertens
P
P
Higgins
L
L
Egelman
E
E
Anderson
D
D
D017103:Bacillus Phages / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D001412:Bacillus subtilis / Q000821:virology; D001499:Bayes Theorem; D002213:Capsid / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D005612:Freeze Drying; D007930:Leucine / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D008961:Models, Structural; D008970:Molecular Weight; D011506:Proteins / Q000302:isolation & purification / Q000648:ultrastructure; D011869:Radioligand Assay; D012588:Scintillation Counting; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014771:Virion / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure
D011506:Proteins; D007930:Leucine