PMID:21705600

Citation	Jakutytė, L, Baptista, C, São-José, C, Daugelavičius, R, Carballido-López, R and Tavares, P (2011) Bacteriophage infection in rod-shaped gram-positive bacteria: evidence for a preferential polar route for phage SPP1 entry in Bacillus subtilis. J. Bacteriol. 193:4893-903
Abstract	Entry into the host bacterial cell is one of the least understood steps in the life cycle of bacteriophages. The different envelopes of Gram-negative and Gram-positive bacteria, with a fluid outer membrane and exposing a thick peptidoglycan wall to the environment respectively, impose distinct challenges for bacteriophage binding and (re)distribution on the bacterial surface. Here, infection of the Gram-positive rod-shaped bacterium Bacillus subtilis by bacteriophage SPP1 was monitored in space and time. We found that SPP1 reversible adsorption occurs preferentially at the cell poles. This initial binding facilitates irreversible adsorption to the SPP1 phage receptor protein YueB, which is encoded by a putative type VII secretion system gene cluster. YueB was found to concentrate at the cell poles and to display a punctate peripheral distribution along the sidewalls of B. subtilis cells. The kinetics of SPP1 DNA entry and replication were visualized during infection. Most of the infecting phages DNA entered and initiated replication near the cell poles. Altogether, our results reveal that the preferentially polar topology of SPP1 receptors on the surface of the host cell determines the site of phage DNA entry and subsequent replication, which occurs in discrete foci.
Links	PubMed PMC3165636 Online version:10.1128/JB.05104-11
Keywords	Bacillus Phages/physiology; Bacillus subtilis/virology; Bacterial Proteins/metabolism; DNA, Viral/metabolism; Protein Binding; Receptors, Virus/metabolism; Virus Attachment; Virus Internalization; Virus Replication
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Significance

Conclusion

The bottom line of this paper is that SPP1 infection of the polar region in Bacillus subtilis is YueB independent even YueB mostly localized in the poles regions. Since in Fig 2d shown that YB886.delta6 also have the same SPPI polar absorption compared to YB886 wild-type.

Questions raised

1)Since WTA is more concentrated at the cell poles, is that the reason why SPP1 preferred the poles?

2) Did the reversible adsorption experiment from 2d done with dilutions as a way to wash off the unbinded phages?

3) Is the Promoter spuc a leaky promoter?

4) There was no negative control (YueB null mutant) for experiment shown in fig 4A

5) Fig7, LacI binding doesn't tell you when everything including CFP is. What is the total CFP? Therefore fig7 isn't informative.

Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status

References

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