RT Journal Article SR Electronic T1 Exploring Genomic Conservation in Actinobacteriophages with Small Genomes JF bioRxiv FD Cold Spring Harbor Laboratory SP 2022.02.05.479200 DO 10.1101/2022.02.05.479200 A1 Vincent Y. Tse A1 Haoyuan Liu A1 Andrew Kapinos A1 Canela Torres A1 Breanna Camille S. Cayabyab A1 Sarah N. Fett A1 Lucy G. Nakashima A1 Mujtahid Rahman A1 Aida S. Vargas A1 Krisanavane Reddi A1 Jordan Moberg Parker A1 Amanda C. Freise YR 2022 UL http://biorxiv.org/content/early/2022/02/06/2022.02.05.479200.abstract AB Actinobacteriophages of a wide range of genome sizes continue to be isolated and characterized, but only a handful of these have atypically small genomes, defined in this work as genome sizes under 20,000 bp. These “small phages” are relatively rare and have received minimal study thus far. Of the actinobacteriophages published in PhagesDB.org, small phages have been isolated on Arthrobacter, Gordonia, Rhodococcus, and Microbacterium hosts. A previous study by Pope et al. showed that Gordonia small phages have similar gene products and amino acid sequences. Here, we set out to further examine relationships between small Gordonia phages as well as small phages that infect other hosts. Of the 3222 sequenced phages listed on PhagesDB, we identified 109 distinctly small phages with genome sizes under 20,000 bp. The majority of the small phages were isolated on Arthrobacter or Microbacterium hosts. Using comparative genomics, we searched for patterns of similarity among 34 cluster-representative small phages. Dot plot comparisons showed that there was more amino acid conservation than nucleotide identity amongst small phages. Gene content similarity (GCS) analysis revealed that the temperate Gordonia phages in Cluster CW share significant GCS values (over 35%) with the lytic Arthrobacter phages in Cluster AN, suggesting that some small phages have a considerable degree of genomic similarity with each other. SplitsTree analyses of shared phams (genes with substantial amino acid identity) supported the complexity of clustering criteria in small phages, given shuffling of genes across phages of different clusters and close relationships despite varied cluster membership. We observed this continuum of phage diversity through Rhodococcus phage RRH1’s closer similarity to phages in Gordonia subcluster CW1 than CW1 is to Gordonia subcluster CW3. Finally, we were able to confirm the presence of conserved phams across not only small Gordonia phages but also within small phages from different clusters and hosts. Studying these genomic trends hidden in small phages allows us to better understand and appreciate the overall diversity of phages.Competing Interest StatementThe authors have declared no competing interest.