FN Archimer Export Format
PT J
TI Cooperation between two modes for DNA replication initiation in the archaeon Thermococcus barophilus
BT 
AF MCTEER, Logan
   Moalic, Yann
   Cueff-Gauchard, Valerie
   Catchpole, Ryan
   Hogrel, Gaëlle
   Lu, Yang
   Laurent, Sebastien
   Hemon, Marie
   Aubé, Johanne
   Leroy, Elodie
   Roussel, Erwan
   Oberto, Jacques
   Flament, Didier
   Dulermo, Remi
AS 1:1;2:2,4;3:1;4:3;5:1;6:1;7:1;8:4;9:1;10:1;11:1;12:3;13:1;14:1;
FF 1:PDG-REM-BEEP-LMEE;2:;3:PDG-REM-BEEP-LMEE;4:;5:;6:PDG-REM-EEP-LMEE;7:PDG-REM-BEEP-LMEE;8:;9:PDG-REM-BEEP-LMEE;10:PDG-REM-BEEP-LMEE;11:PDG-REM-BEEP-LMEE;12:;13:PDG-REM-BEEP-LMEE;14:PDG-REM-BEEP-LMEE;
C1 Univ Brest, Ifremer, CNRS, UMR6197 Biologie et Ecologie des Ecosystèmes marins Profonds (BEEP), Plouzané, France
   LabISEN, Yncréa Ouest, Brest, France
   Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Gif-sur-Yvette, France
   Univ Brest, Ifremer, CNRS, UMR6197 Biologie et Ecologie des Ecosystèmes marins Profonds (BEEP), Plouzané, France
C2 IFREMER, FRANCE
   ISEN, FRANCE
   UNIV PARIS-SACLAY, FRANCE
   UBO, FRANCE
SI BREST
SE PDG-REM-BEEP-LMEE
   PDG-REM-EEP-LMEE
UM BEEP-LM2E
IN DOAJ
IF 6.4
TC 0
UR https://archimer.ifremer.fr/doc/00883/99483/109545.pdf
   https://archimer.ifremer.fr/doc/00883/99483/109546.pdf
   https://archimer.ifremer.fr/doc/00883/99483/109547.pdf
   https://archimer.ifremer.fr/doc/00883/99483/109548.pdf
   https://archimer.ifremer.fr/doc/00883/99483/109549.tif
   https://archimer.ifremer.fr/doc/00883/99483/109550.docx
LA English
DT Article
DE ;DNA replication;homologous recombination;DNA-seq;recombination-dependent replication (RDR);log/stationary phases;recA/Rad51 protein family;Ori;Cdc6
AB The mechanisms underpinning the replication of genomic DNA have recently been challenged in Archaea . Indeed, the lack of origin of replication has no deleterious effect on growth, suggesting that replication initiation relies on homologous recombination. Recombination-dependent replication (RDR) appears to be based on the recombinase RadA, which is of absolute requirement when no initiation origins are detected. The origin of this flexibility in the initiation of replication and the extent to which it is used in nature are yet to be understood. Here, we followed the process of DNA replication throughout the growth stages of Thermococcus barophilus . We combined deep sequencing and genetics to elucidate the dynamics of oriC utilization according to growth phases. We discovered that in T. barophilus , the use of oriC diminishes from the lag to the middle of the log phase, and subsequently increases gradually upon entering the stationary phase. Although oriC demonstrates no indispensability, RadA does exhibit essentiality. Notably, a knockdown mutant strain provides confirmation of the pivotal role of RadA in RDR for the first time. Thus, we demonstrate the existence of a tight combination between oriC utilization and homologous recombination to initiate DNA replication along the growth phases. Overall, this study demonstrates how diverse physiological states can influence the initiation of DNA replication, offering insights into how environmental sensing might impact this fundamental mechanism of life. IMPORTANCE Replication of DNA is highly important in all organisms. It initiates at a specific locus called ori , which serves as the binding site for scaffold proteins—either Cdc6 or DnaA—depending on the domain of life. However, recent studies have shown that the Archaea , Haloferax volcanii and Thermococcus kodakarensis could subsist without ori . Recombination-dependent replication (RDR), via the recombinase RadA, is the mechanism that uses homologous recombination to initiate DNA replication. The extent to which ori ’s use is necessary in natural growth remains to be characterized. In this study, using Thermococcus barophilus , we demonstrated that DNA replication initiation relies on both oriC and RDR throughout its physiological growth, each to varying degrees depending on the phase. Notably, a knockdown RadA mutant confirmed the prominent use of RDR during the log phase. Moreover, the study of ploidy in oriC and radA mutant strains showed that the number of chromosomes per cell is a critical proxy for ensuring proper growth and cell survival. 
PY 2024
PD APR
SO mBio
SN 2150-7511
PU American Society for Microbiology
VL 15
IS 4
DI 10.1128/mbio.03200-23
ID 99483
ER

EF