Genome Sequences of Mycobacteriophages Amgine, Amohnition, Bella96, Cain, DarthP, Hammy, Krueger, LastHope, Peanam, PhelpsODU, Phrank, SirPhilip, Slimphazie, and Unicorn

Anders, Kirk R.; Barekzi, Nazir; Best, Aaron A.; Frederick, Gregory D.; Mavrodi, Dmitri V.; Vazquez, Edwin; Sea-Phages,; Amoh, Nana Yaa A.; Baliraine, Frederick N.; Buchser, William J.; Cast, Thomas P.; Chamberlain, Carmen; Chung, Huimin; D’Angelo, William; Farris, Christian T.; Fernandez-Martinez, Mariceli; Fischman, Haley; Forsyth, Mark H.; Fortier, Anna G.; Gallo, Kara F.; Held, Greta J.; Lomas, Miguel A.; Maldonado-Vazquez, Natalia Y.; Moonsammy, Claudia H.; Namboote, Peace; Paudel, Sudip; Polley, Sarah-Elizabeth M.; Reyes, Gabriella M.; Rubin, Michael R.; Saha, Margaret S.; Stukey, Joseph; Tobias, Tristan; Garlena, Rebecca A.; Stoner, Ty H.; Cresawn, Steven G.; Jacobs-Sera, Deborah; Pope, Welkin H.; Russell, Daniel A.; Hatfull, Graham F.

doi:10.1128/genomea.01202-17

Cited by 3 publications

(2 citation statements)

References 13 publications

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“…Here, we report the results of a genome-wide overexpression screen for Hammy, a temperate siphovirus that infects M. smegmatis. Based on gene content similarity, Hammy is classified as part of Cluster K (Subcluster K6) and is related to Waterfoul and several clinically relevant mycobacteriophages such as ZoeJ (Subcluster K2) (Russell and Hatfull 2016;Anders et al 2017;Dedrick et al 2019b;Guerrero-Bustamante et al 2021). The Hammy genome contains 95 predicted protein-coding genes, only ~35% of which have annotated functions (Figure 1) (Anders et al 2017).…”

Section: Introductionmentioning

confidence: 99%

“…Based on gene content similarity, Hammy is classified as part of Cluster K (Subcluster K6) and is related to Waterfoul and several clinically relevant mycobacteriophages such as ZoeJ (Subcluster K2) (Russell and Hatfull 2016;Anders et al 2017;Dedrick et al 2019b;Guerrero-Bustamante et al 2021). The Hammy genome contains 95 predicted protein-coding genes, only ~35% of which have annotated functions (Figure 1) (Anders et al 2017). Out of 94 Hammy genes evaluated in our systematic screen, a diverse set of 24 genes, representing 25% of the genome, were found to interfere with host growth; 12 of these diverse products caused near complete growth abolition.…”

Section: Introductionmentioning

confidence: 99%

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A genome-wide overexpression screen revealsMycobacterium smegmatisgrowth inhibitors encoded by mycobacteriophage Hammy

Amaya,

Edwards,

Wise

et al. 2023

Preprint

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During infection, bacteriophages produce diverse gene products to overcome bacterial anti-phage defenses, to outcompete other phages, and take over cellular processes. Even in the best-studied model phages, the roles of most phage-encoded gene products are unknown, and the phage population represents a largely untapped reservoir of novel gene functions. Considering the sheer size of this population, experimental screening methods are needed to sort through the enormous collection of available sequences and identify gene products that can modulate bacterial behavior for downstream functional characterization. Here, we describe the construction of a plasmid-based overexpression library of 94 genes encoded by Hammy, a Cluster K mycobacteriophage closely related to those infecting clinically important mycobacteria. The arrayed library was systematically screened in a plate-based cytotoxicity assay, identifying a diverse set of 24 gene products (representing 25% of the Hammy genome) capable of inhibiting growth of the host bacterium Mycobacterium smegmatis. Half of these are related to growth inhibitors previously identified in related phage Waterfoul, supporting their functional conservation; the other genes represent novel additions to the list of known anti-mycobacterial growth inhibitors. This work, conducted as part of the HHMI-supported Science Education Alliance Gene-function Elucidation by a Network of Emerging Scientists (SEA-GENES) project, highlights the value of parallel, comprehensive overexpression screens in exploring genome-wide patterns of phage gene function and novel interactions between phages and their hosts.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A genome-wide overexpression screen revealsMycobacterium smegmatisgrowth inhibitors encoded by mycobacteriophage Hammy

Amaya,

Edwards,

Wise

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

Mycobacteriophages: therapeutic approach for mycobacterial infections

Raman,

Siva Reddy,

Jain

et al. 2024

Drug Discovery Today

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A genome-wide overexpression screen reveals Mycobacterium smegmatis growth inhibitors encoded by mycobacteriophage Hammy

Amaya,

Edwards,

Wise

et al. 2023

G3: Genes, Genomes, Genetics

View full text Add to dashboard Cite

During infection, bacteriophages produce diverse gene products to overcome bacterial antiphage defenses, to outcompete other phages, and to take over cellular processes. Even in the best-studied model phages, the roles of most phage-encoded gene products are unknown, and the phage population represents a largely untapped reservoir of novel gene functions. Considering the sheer size of this population, experimental screening methods are needed to sort through the enormous collection of available sequences and identify gene products that can modulate bacterial behavior for downstream functional characterization. Here, we describe the construction of a plasmid-based overexpression library of 94 genes encoded by Hammy, a Cluster K mycobacteriophage closely related to those infecting clinically important mycobacteria. The arrayed library was systematically screened in a plate-based cytotoxicity assay, identifying a diverse set of 24 gene products (representing ∼25% of the Hammy genome) capable of inhibiting growth of the host bacterium Mycobacterium smegmatis. Half of these are related to growth inhibitors previously identified in related phage Waterfoul, supporting their functional conservation; the other genes represent novel additions to the list of known antimycobacterial growth inhibitors. This work, conducted as part of the HHMI-supported Science Education Alliance Gene-function Exploration by a Network of Emerging Scientists (SEA-GENES) project, highlights the value of parallel, comprehensive overexpression screens in exploring genome-wide patterns of phage gene function and novel interactions between phages and their hosts.

show abstract

Genome Sequences of Mycobacteriophages Amgine, Amohnition, Bella96, Cain, DarthP, Hammy, Krueger, LastHope, Peanam, PhelpsODU, Phrank, SirPhilip, Slimphazie, and Unicorn

Cited by 3 publications

References 13 publications

A genome-wide overexpression screen revealsMycobacterium smegmatisgrowth inhibitors encoded by mycobacteriophage Hammy

A genome-wide overexpression screen revealsMycobacterium smegmatisgrowth inhibitors encoded by mycobacteriophage Hammy

Mycobacteriophages: therapeutic approach for mycobacterial infections

A genome-wide overexpression screen reveals Mycobacterium smegmatis growth inhibitors encoded by mycobacteriophage Hammy

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