A1 Refereed original research article in a scientific journal
The DNA polymerase of bacteriophage YerA41 replicates its T-modified DNA in a primer-independent manner
Authors: Gomez-Raya-Vilanova Miguel V, Leskinen Katarzyna, Bhattacharjee Arnab, Virta Pasi, Rosenqvist Petja, Smith Jake LR, Bayfield Oliver W, Homberger Christina, Kerrinnes Tobias, Vogel Jörg, Pajunen Maria I, Skurnik Mikael
Publisher: OXFORD UNIV PRESS
Publication year: 2022
Journal: Nucleic Acids Research
Journal name in source: NUCLEIC ACIDS RESEARCH
Journal acronym: NUCLEIC ACIDS RES
Volume: 50
Issue: 7
First page : 3985
Last page: 3997
Number of pages: 13
ISSN: 0305-1048
DOI: https://doi.org/10.1093/nar/gkac203
Web address : https://academic.oup.com/nar/article/50/7/3985/6561659
Self-archived copy’s web address: https://research.utu.fi/converis/portal/detail/Publication/175191148
Yersinia phage YerA41 is morphologically similar to jumbo bacteriophages. The isolated genomic material of YerA41 could not be digested by restriction enzymes, and used as a template by conventional DNA polymerases. Nucleoside analysis of the YerA41 genomic material, carried out to find out whether this was due to modified nucleotides, revealed the presence of a ca 1 kDa substitution of thymidine with apparent oligosaccharide character. We identified and purified the phage DNA polymerase (DNAP) that could replicate the YerA41 genomic DNA even without added primers. Cryo-electron microscopy (EM) was used to characterize structural details of the phage particle. The storage capacity of the 131 nm diameter head was calculated to accommodate a significantly longer genome than that of the 145 577 bp genomic DNA of YerA41 determined here. Indeed, cryo-EM revealed, in contrast to the 25 angstrom in other phages, spacings of 33-36 angstrom between shells of the genomic material inside YerA41 heads suggesting that the heavily substituted thymidine increases significantly the spacing of the DNA packaged inside the capsid. In conclusion, YerA41 appears to be an unconventional phage that packages thymidine-modified genomic DNA into its capsids along with its own DNAP that has the ability to replicate the genome.
Downloadable publication This is an electronic reprint of the original article. |  |
