Structural and Functional Studies of a Klebsiella Phage Capsule Depolymerase Tailspike: Mechanistic Insights into Capsular Degradation

Squeglia, Flavia; Maciejewska, Barbara; Łątka, Agnieszka; Ruggiero, Alessia; Briers, Yves; Drulis-Kawa, Zuzanna; Berisio, Rita

doi:10.1016/j.str.2020.04.015

Cited by 46 publications

(57 citation statements)

References 75 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Klebsiella phages KP32, KP34, and KP36 were selected as representatives of well-characterized Klebsiella phage groups. Their RBPs with depolymerase activity were identified and characterized previously (8,9,31), and their RBP architectures differ significantly (16) (see Fig. 2A).…”

Section: Resultsmentioning

confidence: 99%

“…2A). Phage KP32 has two RBPs, with a first RBP connected to the phage tail via an N-terminal anchor domain (1A), while the second RBP (2E) interacts with the first RBP (31). Podovirus KP34 has a single RBP (3E) indirectly attached to the virion via an intermediate anchor protein (3A), and siphovirus KP36 has a single RBP, including an N-terminal anchor domain (4A) for direct attachment to the phage particle.…”

Section: Resultsmentioning

confidence: 99%

“…The construction of chimeric RBPs was based on the former confirmation of the enzymatic activity and specificity of the selected model RBPs with a depolymerase domain (8,9,31). All chimeric RBPs were enzymatically active with a specificity switch corresponding to the predicted enzymatic domain.…”

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Engineering the Modular Receptor-Binding Proteins ofKlebsiellaPhages Switches Their Capsule Serotype Specificity

Łątka

Lemire

Grimon

et al. 2021

mBio

Self Cite

View full text Add to dashboard Cite

The high specificity of bacteriophages is driven by their receptor-binding proteins (RBPs). Many Klebsiella bacteriophages target the capsular exopolysaccharide as the receptor and encode RBPs with depolymerase activity. The modular structure of these RBPs with an N-terminal structural module to attach the RBP to the phage tail, and a C-terminal specificity module for exopolysaccharide degradation, supports horizontal transfer as a major evolutionary driver for Klebsiella phage RBPs. We mimicked this natural evolutionary process by the construction of modular RBP chimeras, exchanging N-terminal structural modules and C-terminal specificity modules. All chimeras strictly follow the capsular serotype specificity of the C-terminal module. Transplanting chimeras with a K11 N-terminal structural RBP module in a Klebsiella phage K11 scaffold results in a capsular serotype switch and corresponding host range modification of the synthetic phages, demonstrating that horizontal transfer of C-terminal specificity modules offers Klebsiella phages an evolutionary highway for rapid adaptation to new capsular serotypes. IMPORTANCE The antimicrobial resistance crisis has rekindled interest in bacteriophage therapy. Phages have been studied over a century as therapeutics to treat bacterial infections, but one of the biggest challenges for the use of phages in therapeutic interventions remains their high specificity. In particular, many Klebsiella phages have a narrow spectrum constrained by the high diversity of exopolysaccharide capsules that shield access to the cells. In this work, we have elaborated how Klebsiella phages deal with this high diversity by exchanging building blocks of their receptor-binding proteins.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Engineering the Modular Receptor-Binding Proteins ofKlebsiellaPhages Switches Their Capsule Serotype Specificity

Łątka

Lemire

Grimon

et al. 2021

mBio

Self Cite

View full text Add to dashboard Cite

show abstract

“…Most of our isolated phages encode at least one gene annotated as a putative “tail-fibre” or “tail-spike” protein (table S2). Intial structural predictions suggested that these proteins adopt beta-helical structures, a common protein architecture of both tail-spike proteins and the capsule depolymerase enzymes that are suggested to have evolved from these purely structural proteins 52–54 . These proteins also contained predicted enzymatic domains, such as the Pectate_lyase_3 domain or Peptidase_S74 domain, which have been identified in other phage encoded depolymerases 55–58 .…”

Section: Resultsmentioning

confidence: 99%

Isolation and characterisation ofKlebsiellaphages for phage therapy

Townsend

Kelly

Gannon

et al. 2020

Preprint

View full text Add to dashboard Cite

Klebsiella is a clinically important pathogen causing a variety of antimicrobial resistant infections, in both community and nosocomial settings, particularly pneumonia, urinary tract infection and septicaemia. We report the successful isolation and characterisation of 30 diverse Klebsiella-infecting phages. The isolated phages are diverse, spanning six different phage families and nine genera. These phages comprise of both lysogenic and lytic lifestyles. Individual Klebsiella phage isolates infected 11 of 18 different Klebsiella capsule types across all six Klebsiella species tested. Our Klebsiella-infecting lytic phages are suitable for phage therapy, based on the criteria that they encode no known toxin or antimicrobial resistance genes. However, none of the characterised phages were able to suppress the growth of Klebsiella for more than 7 hours. This indicates that for successful phage therapy, a mixed cocktail of multiple phages is necessary to treat Klebsiella infections.

show abstract

“…A known characteristic of phage K32 and related phages which belong to Podoviridae is the presence of capsule-polysaccharide specific depolymerase. This enzyme for capsule degradation is required for adsorption to the host cell and subsequent degradation of the capsule in order to expose outer membrane components for phage binding [ 21 , 29 , 30 ]. The distant action of depolymerase on neighboring bacteria may contribute to the observed synergy.…”

Section: Discussionmentioning

confidence: 99%

Kinetic Fingerprinting Links Bacteria-Phage Interactions with Emergent Dynamics: Rapid Depletion of Klebsiella pneumoniae Indicates Phage Synergy

et al. 2020

View full text Add to dashboard Cite

The specific temporal evolution of bacterial and phage population sizes, in particular bacterial depletion and the emergence of a resistant bacterial population, can be seen as a kinetic fingerprint that depends on the manifold interactions of the specific phage–host pair during the course of infection. We have elaborated such a kinetic fingerprint for a human urinary tract Klebsiella pneumoniae isolate and its phage vB_KpnP_Lessing by a modeling approach based on data from in vitro co-culture. We found a faster depletion of the initially sensitive bacterial population than expected from simple mass action kinetics. A possible explanation for the rapid decline of the bacterial population is a synergistic interaction of phages which can be a favorable feature for phage therapies. In addition to this interaction characteristic, analysis of the kinetic fingerprint of this bacteria and phage combination revealed several relevant aspects of their population dynamics: A reduction of the bacterial concentration can be achieved only at high multiplicity of infection whereas bacterial extinction is hardly accomplished. Furthermore the binding affinity of the phage to bacteria is identified as one of the most crucial parameters for the reduction of the bacterial population size. Thus, kinetic fingerprinting can be used to infer phage–host interactions and to explore emergent dynamics which facilitates a rational design of phage therapies.

show abstract

Structural and Functional Studies of a Klebsiella Phage Capsule Depolymerase Tailspike: Mechanistic Insights into Capsular Degradation

Cited by 46 publications

References 75 publications

Engineering the Modular Receptor-Binding Proteins ofKlebsiellaPhages Switches Their Capsule Serotype Specificity

Engineering the Modular Receptor-Binding Proteins ofKlebsiellaPhages Switches Their Capsule Serotype Specificity

Isolation and characterisation ofKlebsiellaphages for phage therapy

Kinetic Fingerprinting Links Bacteria-Phage Interactions with Emergent Dynamics: Rapid Depletion of Klebsiella pneumoniae Indicates Phage Synergy

Contact Info

Product

Resources

About