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The terminase motors of bacteriophages have been shown to be among thestrongest active machines in the biomolecular world, being able to packageseveral tens of kilobase pairs of viral genome into a capsid within minutes.Yet these motors are hindered at the end of the packaging process by theprogressive build-up of a force resisting packaging associated with alreadypackaged DNA. In this experimental work, we raise the issue of what sets theupper limit on the length of the genome that can be packaged by the terminasemotor of phage {\lambda} and still yield infectious virions, and the conditionsunder which this can be efficiently performed. Using a packaging strategydeveloped in our laboratory of building phage {\lambda} from scratch, togetherwith plaque assay monitoring, we have been able to show that the terminasemotor of phage {\lambda} is able to produce infectious particles with up to110% of the wild-type (WT) {\lambda}-DNA length. However, the phage productionrate, and thus the infectivity, decreased exponentially with increasing DNAlength, and was a factor of 103 lower for the 110% {\lambda}-DNA phage.Interestingly, our in vitro strategy was still efficient in fully packagingphages with DNA lengths as high as 114% of the WT length, but these viruseswere unable to infect bacterial cells efficiently. Further, we demonstratedthat the phage production rate is modulated by the presence of multivalentionic species. The biological consequences of these finding are discussed.

Challenging packaging limits and infectivity of phage {\lambda}