PT  - JOURNAL ARTICLE
AU  - David Lukac
AU  - Zuzana Machacova
AU  - Pavel Moudry
TI  - Emetine blocks DNA replication via proteosynthesis inhibition not by targeting Okazaki fragments
AID  - 10.26508/lsa.202201560
DP  - 2022 Dec 01
TA  - Life Science Alliance
PG  - e202201560
VI  - 5
IP  - 12
4099  - https://www.life-science-alliance.org/content/5/12/e202201560.short
4100  - https://www.life-science-alliance.org/content/5/12/e202201560.full
SO  - Life Sci. Alliance2022 Dec 01; 5
AB  - DNA synthesis of the leading and lagging strands works independently and cells tolerate single-stranded DNA generated during strand uncoupling if it is protected by RPA molecules. Natural alkaloid emetine is used as a specific inhibitor of lagging strand synthesis, uncoupling leading and lagging strand replication. Here, by analysis of lagging strand synthesis inhibitors, we show that despite emetine completely inhibiting DNA replication: it does not induce the generation of single-stranded DNA and chromatin-bound RPA32 (CB-RPA32). In line with this, emetine does not activate the replication checkpoint nor DNA damage response. Emetine is also an inhibitor of proteosynthesis and ongoing proteosynthesis is essential for the accurate replication of DNA. Mechanistically, we demonstrate that the acute block of proteosynthesis by emetine temporally precedes its effects on DNA replication. Thus, our results are consistent with the hypothesis that emetine affects DNA replication by proteosynthesis inhibition. Emetine and mild POLA1 inhibition prevent S-phase poly(ADP-ribosyl)ation. Collectively, our study reveals that emetine is not a specific lagging strand synthesis inhibitor with implications for its use in molecular biology.