Experimental test of connector rotation during DNA packaging into bacteriophage phi29 capsids.
PLoS Biol
; 5(3): e59, 2007 Mar.
Article
em En
| MEDLINE
| ID: mdl-17311473
ABSTRACT
The bacteriophage phi29 generates large forces to compact its double-stranded DNA genome into a protein capsid by means of a portal motor complex. Several mechanical models for the generation of these high forces by the motor complex predict coupling of DNA translocation to rotation of the head-tail connector dodecamer. Putative connector rotation is investigated here by combining the methods of single-molecule force spectroscopy with polarization-sensitive single-molecule fluorescence. In our experiment, we observe motor function in several packaging complexes in parallel using video microscopy of bead position in a magnetic trap. At the same time, we follow the orientation of single fluorophores attached to the portal motor connector. From our data, we can exclude connector rotation with greater than 99% probability and therefore answer a long-standing mechanistic question.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA Viral
/
Capsídeo
/
Fagos Bacilares
/
Empacotamento do DNA
Tipo de estudo:
Prognostic_studies
Idioma:
En
Revista:
PLoS Biol
Assunto da revista:
BIOLOGIA
Ano de publicação:
2007
Tipo de documento:
Article
País de afiliação:
Alemanha