A group II intron-encoded protein interacts with the cellular replicative machinery through the ß-sliding clamp

García-Rodríguez, Fernando M. ; Neira, José L. (Universidad de Zaragoza) ; Marcia, Marco ; Molina-Sánchez, María D. ; Toro, Nicolás

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 ß-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 ß-sliding clamp of the DNA replication machinery, favouring reverse splicing into the transient ssDNA at DNA replication forks.

Idioma: Inglés

DOI: 10.1093/nar/gkz468

Año: 2019

Publicado en: Nucleic Acids Research 47, 14 (2019), 7605-7617
ISSN: 0305-1048

Factor impacto JCR: 11.501 (2019)

Categ. JCR: BIOCHEMISTRY & MOLECULAR BIOLOGY rank: 15 / 297 = 0.051 (2019) - Q1 - T1

Factor impacto SCIMAGO: 8.907 - Genetics (Q1)

Financiación: info:eu-repo/grantAgreement/ES/MINECO-FEDER/BIO2014-51953-P
Financiación: info:eu-repo/grantAgreement/ES/MINECO-FEDER/BIO2017-82244-P

Tipo y forma: Article (Published version)

Área (Departamento): Área Lenguajes y Sistemas Inf. (Dpto. Informát.Ingenie.Sistms.)