Real-time observations of single bacteriophage λ DNA ejections in vitro

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 104; no. 37; pp. 14652 - 14657
• Main Authors: Grayson, Paul, Han, Lin, Winther, Tabita, Phillips, Rob
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 11.09.2007; National Acad Sciences
• Series: From the Cover
• Subjects: Bacteria; Bacteriophage lambda - genetics; Bacteriophage lambda - physiology; Bacteriophages; Biological Sciences; Biological Transport, Active - physiology; Buffers; Capsid; Cations, Divalent - chemistry; DNA; DNA, Viral - chemistry; DNA, Viral - physiology; Dyes; Escherichia coli - metabolism; Escherichia coli - virology; Fluorescence; fluorescence microscopy; friction; genome; Genome, Viral; Genomes; head; In Vitro Techniques; ions; magnesium; Magnesium - chemistry; Microscopy; Microscopy, Fluorescence; Osmolar Concentration; Phage l; Phage T5; Physics; Pressure; sodium; Speed; Trajectories
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.0703274104

The physical, chemical, and structural features of bacteriophage genome release have been the subject of much recent attention. Many theoretical and experimental studies have centered on the internal forces driving the ejection process. Recently, Mangenot et al. [Mangenot S, Hochrein M, Rädler J, Letellier L (2005) Curr Biol 15:430-435.] reported fluorescence microscopy of phage T5 ejections, which proceeded stepwise between DNA nicks, reaching a translocation speed of 75 kbp/s or higher. It is still unknown how high the speed actually is. This paper reports real-time measurements of ejection from phage λ, revealing how the speed depends on key physical parameters such as genome length and ionic state of the buffer. Except for a pause before DNA is finally released, the entire 48.5-kbp genome is translocated in [almost equal to]1.5 s without interruption, reaching a speed of 60 kbp/s. The process gives insights particularly into the effects of two parameters: a shorter genome length results in lower speed but a shorter total time, and the presence of divalent magnesium ions (replacing sodium) reduces the pressure, increasing ejection time to 8-11 s. Pressure caused by DNA-DNA interactions within the head affects the initiation of ejection, but the close packing is also the dominant source of friction: more tightly packed phages initiate ejection earlier, but with a lower initial speed. The details of ejection revealed in this study are probably generic features of DNA translocation in bacteriophages and have implications for the dynamics of DNA in other biological systems.