Next Article in Journal
Demethylation of Non-CpG Sites in DNA Is Initiated by TET2 5-Methylcytosine Dioxygenase
Previous Article in Journal
The Pre-Metaphase Stretch: A Re-Examination
Article

Ligase A and RNase HI Participate in Completing Replication on the Chromosome in Escherichia coli

1
Department of Microbiology, Cornell University, Ithaca, NY 14853, USA
2
Department of Biology, Portland State University, Portland, OR 97201, USA
*
Author to whom correspondence should be addressed.
Academic Editors: Rodrigo Bermejo and Darren Griffin
Received: 1 July 2021 / Revised: 12 August 2021 / Accepted: 30 August 2021 / Published: 3 September 2021
In Escherichia coli, several enzymes have been identified that participate in completing replication on the chromosome, including RecG, SbcCD, ExoI, and RecBCD. However, other enzymes are likely to be involved and the precise enzymatic mechanism by which this reaction occurs remains unknown. Two steps predicted to be necessary to complete replication are removal of Okazaki RNA fragments and ligation of the nascent strands at convergent replication forks. E. coli encodes two RNases that remove RNA-DNA hybrids, rnhA and rnhB, as well as two ligases, ligA and ligB. Here, we used replication profiling to show that rnhA and ligA, encoding RNase HI and Ligase A, participate in the completion reaction. Deletion of rnhA impaired the ability to complete replication and resulted in over-replication in the terminus region. It additionally suppressed initiation events from oriC, suggesting a role for the enzyme in oriC-dependent initiation, as has been suggested previously. We also show that a temperature-sensitive mutation in Ligase A led to over-replication at sites where replication completes, and that degradation at these sites occurred upon shifting to the nonpermissive temperature. Deletion of rnhB or ligB did not affect the growth or profile of replication on the genome. View Full-Text
Keywords: ligase; RNase H; completion of DNA replication ligase; RNase H; completion of DNA replication
Show Figures

Graphical abstract

MDPI and ACS Style

Wendel, B.M.; Hernandez, A.J.; Courcelle, C.T.; Courcelle, J. Ligase A and RNase HI Participate in Completing Replication on the Chromosome in Escherichia coli. DNA 2021, 1, 13-25. https://0-doi-org.brum.beds.ac.uk/10.3390/dna1010003

AMA Style

Wendel BM, Hernandez AJ, Courcelle CT, Courcelle J. Ligase A and RNase HI Participate in Completing Replication on the Chromosome in Escherichia coli. DNA. 2021; 1(1):13-25. https://0-doi-org.brum.beds.ac.uk/10.3390/dna1010003

Chicago/Turabian Style

Wendel, Brian M., Adrian J. Hernandez, Charmain T. Courcelle, and Justin Courcelle. 2021. "Ligase A and RNase HI Participate in Completing Replication on the Chromosome in Escherichia coli" DNA 1, no. 1: 13-25. https://0-doi-org.brum.beds.ac.uk/10.3390/dna1010003

Find Other Styles

Article Access Map by Country/Region

1
Back to TopTop