AUTHOR=Oliveira Paiva Ana M. , van Eijk Erika , Friggen Annemieke H. , Weigel Christoph , Smits Wiep Klaas 

TITLE=Identification of the Unwinding Region in the Clostridioides difficile Chromosomal Origin of Replication

JOURNAL=Frontiers in Microbiology

VOLUME=11

YEAR=2020

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2020.581401

DOI=10.3389/fmicb.2020.581401

ISSN=1664-302X

ABSTRACT=<p>Faithful DNA replication is crucial for viability of cells across all kingdoms. Targeting DNA replication is a viable strategy for inhibition of bacterial pathogens. <italic>Clostridioides difficile</italic> is an important enteropathogen that causes potentially fatal intestinal inflammation. Knowledge about DNA replication in this organism is limited and no data is available on the very first steps of DNA replication. Here, we use a combination of <italic>in silico</italic> predictions and <italic>in vitro</italic> experiments to demonstrate that <italic>C. difficile</italic> employs a bipartite origin of replication that shows DnaA-dependent melting at <italic>oriC2</italic>, located in the <italic>dnaA</italic>-<italic>dnaN</italic> intergenic region. Analysis of putative origins of replication in different clostridia suggests that the main features of the origin architecture are conserved. This study is the first to characterize aspects of the origin region of <italic>C. difficile</italic> and contributes to our understanding of the initiation of DNA replication in clostridia.</p>