Data/hora: 01/06/2024 06:34:24

Provedor de dados: | 5
País: | France
Título: | In vitro reconstitution of RNA primer removal in Archaea reveals the existence of two pathways
Autores: | Henneke, Ghislaine.
Data: | 2012-10
Ano: | 2012
Palavras-chave: | Archaea; DNA ligase; DNA polymerase; Nuclease; Okazaki fragment.
Resumo: | Using model DNA substrates and purified recombinant proteins from Pyrococcus abyssi. I have reconstituted the enzymatic reactions involved in RNA primer elimination in vitro. In my dual-labelled system, polymerase D performed efficient strand displacement DNA synthesis, generating 5'-RNA flaps which were subsequently released by Fen1, before ligation by Lig1. In this pathway, the initial cleavage event by RNase HII facilitated RNA primer removal of Okazaki fragments. In addition, I have shown that polymerase B was able to displace downstream DNA strands with a single ribonucleotide at the 5'-end, a product resulting from a single cut in the RNA initiator by RNase HII. After RNA elimination, the combined activities of strand displacement DNA synthesis by polymerase B and flap cleavage by Fen1 provided a nicked substrate for ligation by Lig1. The unique specificities of Okazaki fragment maturation enzymes and replicative DNA polymerases strongly support the existence of two pathways in the resolution of RNA fragments.
Tipo: | Text
Idioma: | Inglês

Identificador:

http://archimer.ifremer.fr/doc/00107/21841/20923.pdf

DOI:10.1042/BJ20120959

Editor: | Portland Press Ltd
Relação: | http://archimer.ifremer.fr/doc/00107/21841/
Formato: | application/pdf
Fonte: | Biochemical Journal (0264-6021) (Portland Press Ltd), 2012-10 , Vol. 447 , P. 271-280
Fonte: | The Authors Journal compilation, Biochemical Society