Genome-Wide Mapping of Autonomously Replicating Sequences in the Marine Diatom Phaeodactylum tricornutum

Mar Biotechnol (NY). 2024 Nov 28;27(1):14. doi: 10.1007/s10126-024-10390-0.

Abstract

Autonomously replicating sequences (ARSs) are important accessories in episomal vectors that allow them to be replicated and stably maintained within transformants. Despite their importance, no information on ARSs in diatoms has been reported. Therefore, we attempted to identify ARS candidates in the model diatom, Phaeodactylum tricornutum, via chromatin immunoprecipitation sequencing. In this study, subunits of the origin recognition complex (ORC), ORC2 and ORC4, were used to screen for ARS candidates. ORC2 and ORC4 bound to 355 sites on the P. tricornutum genome, of which 69 were constantly screened after multiple attempts. The screened ARS candidates had an AT-richness of approximately 50% (44.39-52.92%) and did not have conserved sequences. In addition, ARS candidates were distributed randomly but had a dense distribution pattern at several sites. Their positions tended to overlap with those of the genetic region (73.91%). Compared to the ARSs of several other eukaryotic organisms, the characteristics of the screened ARS candidates are complex. Thus, our findings suggest that the diatom has a distinct and unique native ARSs.

Keywords: Phaeodactylum tricornutum; Autonomously replicating sequences; Chromatin immunoprecipitation sequencing; Diatom; Origin recognition complex.

MeSH terms

  • Chromatin Immunoprecipitation Sequencing
  • Chromosome Mapping
  • DNA Replication
  • Diatoms* / genetics
  • Diatoms* / metabolism
  • Genome
  • Origin Recognition Complex / genetics
  • Origin Recognition Complex / metabolism

Substances

  • Origin Recognition Complex