The DNA polymerase of bacteriophage YerA41 replicates its T-modified DNA in a primer-independent manner
Bayfield Oliver W; Kerrinnes Tobias; Vogel Jörg; Virta Pasi; Leskinen Katarzyna; Gomez-Raya-Vilanova Miguel V; Smith Jake LR; Rosenqvist Petja; Skurnik Mikael; Homberger Christina; Pajunen Maria I; Bhattacharjee Arnab
The DNA polymerase of bacteriophage YerA41 replicates its T-modified DNA in a primer-independent manner
Bayfield Oliver W
Kerrinnes Tobias
Vogel Jörg
Virta Pasi
Leskinen Katarzyna
Gomez-Raya-Vilanova Miguel V
Smith Jake LR
Rosenqvist Petja
Skurnik Mikael
Homberger Christina
Pajunen Maria I
Bhattacharjee Arnab
OXFORD UNIV PRESS
Julkaisun pysyvä osoite on:
https://urn.fi/URN:NBN:fi-fe2022081154268
https://urn.fi/URN:NBN:fi-fe2022081154268
Tiivistelmä
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.
Kokoelmat
- Rinnakkaistallenteet [19207]