DNA knots reveal a chiral organization of DNA in phage capsids.
Icosahedral bacteriophages pack their double-stranded DNA genomes to near-crystalline density and achieve one of the highest levels of DNA condensation found in nature. Despite numerous studies, some essential properties of the packaging geometry of the DNA inside the phage capsid are still unknown. We present a different approach to the problems of randomness and chirality of the packed DNA. We recently showed that most DNA molecules extracted from bacteriophage P4 are highly knotted because of the cyclization of the linear DNA molecule confined in the phage capsid. Here, we show that these knots provide information about the global arrangement of the DNA inside the capsid. First, we analyze the distribution of the viral DNA knots by high-resolution gel electrophoresis. Next, we perform Monte Carlo computer simulations of random knotting for freely jointed polygons confined to spherical volumes. Comparison of the knot distributions obtained by both techniques produces a topological proof of nonrandom packaging of the viral DNA. Moreover, our simulations show that the scarcity of the achiral knot 4(1) and the predominance of the torus knot 5(1) over the twist knot 5(2) observed in the viral distribution of DNA knots cannot be obtained by confinement alone but must include writhe bias in the conformation sampling. These results indicate that the packaging geometry of the DNA inside the viral capsid is writhe-directed.
Arsuaga
Javier
J
Departments of Mathematics, Molecular and Cell Biology, and Physics, University of California, Berkeley, CA 94720, USA. jrbbmc@cid.csic.es
Vazquez
Mariel
M
McGuirk
Paul
P
Trigueros
Sonia
S
Sumners
De Witt
D
Roca
Joaquim
J
D001435:Bacteriophages / Q000378:metabolism*; D002213:Capsid / Q000737:chemistry*; D003198:Computer Simulation; D004270:DNA, Circular; D004278:DNA, Superhelical; D004279:DNA, Viral / Q000737:chemistry*; D004587:Electrophoresis, Agar Gel; D005814:Genes, Viral; D009010:Monte Carlo Method; D009690:Nucleic Acid Conformation; D013237:Stereoisomerism
D004270:DNA, Circular; D004278:DNA, Superhelical; D004279:DNA, Viral