Handcuffing reversal is facilitated by proteases and replication initiator monomers

Katarzyna Bury , Katarzyna Węgrzyn , Igor Konieczny


Specific nucleoprotein complexes are formed strictly to prevent over-initiation of DNA replication. An example of those is the so-called handcuff complex, in which two plasmid molecules are coupled together with plasmid-encoded replication initiation protein (Rep). In this work, we elucidate the mechanism of the handcuff complex disruption. In vitro tests, including dissociation progress analysis, demonstrate that the dimeric variants of plasmid RK2 replication initiation protein TrfA are involved in assembling the plasmid handcuff complex which, as we found, reveals high stability. Particular proteases, namely Lon and ClpAP, disrupt the handcuff by degrading TrfA, thus affecting plasmid stability. Moreover, our data demonstrate that TrfA monomers are able to dissociate handcuffed plasmid molecules. Those monomers displace TrfA molecules, which are involved in handcuff formation, and through interaction with the uncoupled plasmid replication origins they re-initiate DNA synthesis. We discuss the relevance of both Rep monomers and host proteases for plasmid maintenance under vegetative and stress conditions.
Author Katarzyna Bury (IFB/IB/DMCB)
Katarzyna Bury,,
- Department of Molecular and Cellular Biology
, Katarzyna Węgrzyn (IFB/IB/DMCB)
Katarzyna Węgrzyn,,
- Department of Molecular and Cellular Biology
, Igor Konieczny (IFB/IB/DMCB)
Igor Konieczny,,
- Department of Molecular and Cellular Biology
Other language title versions
Journal seriesNucleic Acids Research, ISSN 0305-1048, (A 40 pkt)
Issue year2017
Publication size in sheets0.65
ASJC Classification1311 Genetics
URL https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/gkx166
Languageen angielski
LicenseJournal (articles only); published final; Uznanie Autorstwa - Użycie Niekomercyjne (CC-BY-NC); with publication
Score (nominal)40
Score sourcejournalList
ScoreMinisterial score = 40.0, 30-06-2020, ArticleFromJournal
Publication indicators WoS Citations = 3.000; Scopus SNIP (Source Normalised Impact per Paper): 2016 = 2.657; WoS Impact Factor: 2017 = 11.561 (2) - 2017=10.235 (5)
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