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The linker region of LINEs modulates DNA cleavage and DNA polymerization.
Pradhan, Monika; Govindaraju, Aruna; Jagdish, Athena; Christensen, Shawn M.
Afiliação
  • Pradhan M; Department of Biology, University of Texas at Arlington, Arlington, TX, 76019, USA.
  • Govindaraju A; Department of Biology, University of Texas at Arlington, Arlington, TX, 76019, USA.
  • Jagdish A; Department of Biology, University of Texas at Arlington, Arlington, TX, 76019, USA.
  • Christensen SM; Department of Biology, University of Texas at Arlington, Arlington, TX, 76019, USA. Electronic address: shawnc@uta.edu.
Anal Biochem ; 603: 113809, 2020 08 15.
Article em En | MEDLINE | ID: mdl-32511965
ABSTRACT
Long interspersed elements (LINEs) replicate by target primed reverse transcription (TPRT). Insertion involves two half reactions. Each half reaction involves DNA cleavage followed by DNA synthesis. The linker region, located just beyond the reverse transcriptase in the LINE open reading frame, contains a set of predicted helices that may form an α-finger, followed by a gag-like zinc-knuckle. Point mutations of moderately conserved amino-acid residues in the presumptive α-finger severely impair the DNA endonuclease and reverse transcriptase activities of the integration reaction during both half reactions. Mutations in the gag-like zinc-knuckle also impair DNA cleavage and DNA synthesis in some instances. Mutations in core residues that presumably disrupt the protein structure of the presumptive α-finger and the gag-like zinc-knuckle lead to a promiscuous DNA endonuclease and protein-nucleic acid complexes that get stuck in the well during analysis. The linker region appears to function as a protein, DNA, and RNA conformational switching area. The linker is used to properly position nucleic acid substrates into the active sites of the reverse transcriptase and of the DNA endonuclease.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: DNA / Elementos Nucleotídeos Longos e Dispersos Idioma: En Ano de publicação: 2020 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: DNA / Elementos Nucleotídeos Longos e Dispersos Idioma: En Ano de publicação: 2020 Tipo de documento: Article