A group II intron-encoded protein interacts with the cellular replicative machinery through the beta-sliding clamp
Por:
Garcia-Rodriguez, F, Neira, J, Marcia, M, Molina-Sanchez, M and Toro, N
Publicada:
22 ago 2019
Resumen:
Group II introns are self-splicing mobile genetic retroelements. The spliced intron RNA and the intron-encoded protein (IEP) form ribonucleoprotein particles (RNPs) that recognize and invade specific DNA target sites. The IEP is a reverse transcriptase/maturase that may bear a C-terminal endonuclease domain enabling the RNP to cleave the target DNA strand to prime reverse transcription. However, some mobile introns, such as RmInt1, lack the En domain but nevertheless retrohome efficiently to transient single-stranded DNA target sites at a DNA replication fork. Their mobility is associated with host DNA replication, and they use the nascent lagging strand as a primer for reverse transcription. We searched for proteins that interact with RmInt1 RNPs and direct these RNPs to the DNA replication fork. Co-immunoprecipitation assays suggested that DnaN (the beta-sliding clamp), a component of DNA polymerase III, interacts with the protein component of the RmInt1 RNP. Pulldown assays, far-western blots and biolayer interferometry supported this interaction. Peptide binding assays also identified a putative DnaN-interacting motif in the RmInt1 IEP structurally conserved in group II intron IEPs. Our results suggest that intron RNP interacts with the beta-sliding clamp of the DNA replication machinery, favouring reverse splicing into the transient ssDNA at DNA replication forks.
Filiaciones:
Garcia-Rodriguez, F:
CSIC, Struct Dynam & Funct Rhizobacterial Genomes, Dept Soil Microbiol & Symbiot Syst, Estn Expt Zaidin,Grp Ecol Genet Rizosfera, C Prof Albareda 1, E-18008 Granada, Spain
:
Univ Miguel Hernandez, Inst Biol & Mol Celular, Elche 03202, Alicante, Spain
Univ Zaragoza, Joint Units IQFR CSIC BIFI, Inst Biocomp & Fis Sistemas Complejos, E-50009 Zaragoza, Spain
Univ Zaragoza, GBsC CSIC BIFI, E-50009 Zaragoza, Spain
Marcia, M:
EMBL, Grenoble Outstn, 71 Ave Martyrs, F-38042 Grenoble, France
Molina-Sanchez, M:
CSIC, Struct Dynam & Funct Rhizobacterial Genomes, Dept Soil Microbiol & Symbiot Syst, Estn Expt Zaidin,Grp Ecol Genet Rizosfera, C Prof Albareda 1, E-18008 Granada, Spain
Toro, N:
CSIC, Struct Dynam & Funct Rhizobacterial Genomes, Dept Soil Microbiol & Symbiot Syst, Estn Expt Zaidin,Grp Ecol Genet Rizosfera, C Prof Albareda 1, E-18008 Granada, Spain
gold, Green Published, Green Submitted
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