Protocol

Optical Tweezers

Volume 1486 of the series Methods in Molecular Biology pp 343-355

Date:

Deciphering the Molecular Mechanism of the Bacteriophage φ29 DNA Packaging Motor

  • Shixin LiuAffiliated withThe Rockefeller University Email author 
  • , Sara TafoyaAffiliated withUniversity of California, Berkeley
  • , Carlos BustamanteAffiliated withUniversity of California and Howard Hughes Medical Institute, Berkeley Email author 

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Abstract

The past decade has seen an explosion in the use of single-molecule approaches to study complex biological processes. One such approach—optical trapping—is particularly well suited for investigating molecular motors, a diverse group of macromolecular complexes that convert chemical energy into mechanical work, thus playing key roles in virtually every aspect of cellular life. Here we describe how to use high-resolution optical tweezers to investigate the mechanism of the bacteriophage φ29 DNA packaging motor, a ring-shaped ATPase responsible for genome packing during viral assembly. This system illustrates how to use single-molecule techniques to uncover novel, often unexpected, principles of motor operation.

Key words

Single-molecule manipulation Optical tweezers Viral DNA packaging Molecular motor Ring ATPase