title: Klebsiella Phage ΦK64-1 Encodes Multiple Depolymerases for Multiple Host Capsular Types.
abstract: The genome of the multihost bacteriophage ΦK64-1, capable of infectingKlebsiellacapsular types K1, K11, K21, K25, K30, K35, K64, and K69, as well as new capsular types KN4 and KN5, was analyzed and revealed that 11 genes (S1-1,S1-2,S1-3,S2-1,S2-2,S2-3,S2-4,S2-5,S2-6,S2-7, andS2-8) encode proteins with amino acid sequence similarity to tail fibers/spikes or lyases.S2-5previously was shown to encode a K64 capsule depolymerase (K64dep). Specific capsule-degrading activities of an additional eight putative capsule depolymerases (S2-4 against K1, S1-1 against K11, S1-3 against K21, S2-2 against K25, S2-6 against K30/K69, S2-3 against K35, S1-2 against KN4, and S2-1 against KN5) was demonstrated by expression and purification of the recombinant proteins. Consistent with the capsular type-specific depolymerization activity of these gene products, phage mutants ofS1-2,S2-2,S2-3, orS2-6lost infectivity for KN4, K25, K35, or K30/K69, respectively, indicating that capsule depolymerase is crucial for infecting specific hosts. In conclusion, we identified nine functional capsule depolymerase-encoding genes in a bacteriophage and correlated activities of the gene products to all ten hosts of this phage, providing an example of type-specific host infection mechanisms in a multihost bacteriophage.IMPORTANCEWe currently identified eight novel capsule depolymerases in a multihostKlebsiellabacteriophage and correlated the activities of the gene products to all hosts of this phage, providing an example of carriage of multiple depolymerases in a phage with a wide capsular type host spectrum. Moreover, we also established a recombineering system for modification ofKlebsiellabacteriophage genomes and demonstrated the importance of capsule depolymerase for infecting specific hosts. Based on the powerful tool for modification of phage genome, further studies can be conducted to improve the understanding of mechanistic details ofKlebsiellaphage infection. Furthermore, the newly identified capsule depolymerases will be of great value for applications in capsular typing.
authors.affiliation: Department of Microbiology, School of Medicine, China Medical University, Taichung, Taiwan.
authors.forename: Yi-Jiun
authors.initials: YJ
authors.lastname: Pan
authors.affiliation: Department of Microbiology, National Taiwan University College of Medicine, Taipei, Taiwan.
authors.forename: Tzu-Lung
authors.initials: TL
authors.lastname: Lin
authors.affiliation: Department of Microbiology, National Taiwan University College of Medicine, Taipei, Taiwan.
authors.forename: Ching-Ching
authors.initials: CC
authors.lastname: Chen
authors.affiliation: Department of Microbiology, National Taiwan University College of Medicine, Taipei, Taiwan.
authors.forename: Yun-Ting
authors.initials: YT
authors.lastname: Tsai
authors.affiliation: Department of Microbiology, National Taiwan University College of Medicine, Taipei, Taiwan.
authors.forename: Yi-Hsiang
authors.initials: YH
authors.lastname: Cheng
authors.affiliation: Department of Microbiology, National Taiwan University College of Medicine, Taipei, Taiwan.
authors.forename: Yi-Yin
authors.initials: YY
authors.lastname: Chen
authors.affiliation: Department of Microbiology, National Taiwan University College of Medicine, Taipei, Taiwan.
authors.forename: Pei-Fang
authors.initials: PF
authors.lastname: Hsieh
authors.affiliation: Division of Infectious Diseases, Department of Medicine, Taipei Veterans General Hospital, Taipei, Taiwan.
authors.forename: Yi-Tsung
authors.initials: YT
authors.lastname: Lin
authors.affiliation: Department of Microbiology, National Taiwan University College of Medicine, Taipei, Taiwan wangjt@ntu.edu.tw.|Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan.
authors.forename: Jin-Town
authors.initials: JT
authors.lastname: Wang
mesh_terms: D016667:Bacterial Capsules / Q000378:metabolism*; D001435:Bacteriophages / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular; D015870:Gene Expression; D006867:Hydrolases / Q000235:genetics* / Q000378:metabolism*; D007709:Klebsiella / Q000821:virology*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D006867:Hydrolases
keywords: Klebsiella; bacteriophage; capsular type; capsule depolymerase; multiple host