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Fig. 6 | Mobile DNA

Fig. 6

From: LINE-1 ORF1p does not determine substrate preference for human/orangutan SVA and gibbon LAVA

Fig. 6

L1 ORF1p does not confer substrate preference for human SVA versus orangutan SVA versus gibbon LAVA. a Schematic representation of the L1 constructs used. Expression of codon-optimized species-specific ORF1 and human (L1PA1) ORF2 is driven by a CMV promoter. Reporter constructs used to assay for L1 mobilization in cis carry the mneoI reporter cassette; expression constructs for SVA/LAVA mobilization in trans a polyadenylation signal directly following ORF2. b Mobilization of the L1 ORF1p chimeras in cis. Retrotransposition rates +/− SEM are shown relative to the human (L1PA1) construct (100%). Average colony counts are given on top of each column (n = 4). c Mobilization of human and orangutan SVAs and gibbon LAVA in trans. Retrotransposition rates +/− SEM are shown relative to the combination of human L1 – human SVA (100%). Average colony counts are given on top of each column (n = 6). (d) Mutation of PA3 ORF1p amino acids 35 and 172 does not rescue mobilization capacity for SVA in trans. L1PA1, the chimeric L1PA3-PA1 and two mutants thereof were tested in combination with the mneoM-tagged human H8_43 SVA element. Retrotransposition rates +/− SEM are shown relative to the human (L1PA1) construct (100%, n = 4). The insets show Northern blot analyses of the RNAs transcribed from the respective L1 reporter (b) or expression (c) constructs

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