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. 2015 Mar 4;7(3):939–968. doi: 10.3390/v7030939

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© 2015 by the authors; licensee MDPI, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

HML-2 Promoter Expression in Tera-1 Cells. (A) Comparison of the relative transcript expression level (FPKM; black) for a provirus and its corresponding relative luciferase expression level in Tera-1 cells transfected with a vector containing a luciferase reporter gene downstream of the indicated proviral 5’ LTR. LTR activity is expressed as relative light units (RLU; gray) normalized to a control construct with a Renilla luciferase gene driven by an SV40 promoter. The relative promoter activities of the LTR Hs located 551 bp upstream from the 22q11.23 provirus, the 5’ LTR 5B of the 22q11.23 provirus and the 5’ LTR Hs of six other expressed proviruses in Tera-1 cells are shown. (B) Schematic of the 22q11.23 LTR Hs, showing the U3, R and U5 regions. Predicted transcriptional start sites are indicated with black arrows and nucleotide position. Colored boxes indicate previously described promoter element motifs [62,63,64]. Lines below the LTR diagram indicate the regions included in each truncated LTR construct, and numbers to the right of each line indicate the nucleotide position at which the LTR was truncated. GA, GA rich motif (nt 379–386, sequence GGGAAGGG); E, enhancer box (nt 465–476, sequence TTGCAGTTGAGA; nt 485–496, sequence AGGCATCTGTCT; nt 832–843, sequence CTCCATATGCTG); GC, GC rich motif nt 759–763, (sequence CCCCC; nt 602–606, sequence GGCGG); TATA, TATA box (nt 790–797, sequence AATAAATA); Inr, initiator element (nt 807–812, sequence CTCAGA). Cartoon is not drawn to scale. (C) Relative promoter expression levels of truncated 22q11.23 LTR Hs constructs in Tera-1 cells (Kruskal-Wallis, * p < 0.05, ** p < 0.01). All luciferase experiments were conducted in triplicate and are shown as mean ± standard deviation. (D) Schematic of promoter motifs found in the 22q11.21 provirus 5’ LTR Hs, the 22q11.23 LTR Hs and 22q11.23 provirus 5’ LTR 5B. Crossed out boxes indicate presence of a mutation in the motif as compared to the canonical sequence. Cartoon is not drawn to scale.

HML-2 Promoter Expression in Tera-1 Cells. (A) Comparison of the relative transcript expression level (FPKM; black) for a provirus and its corresponding relative luciferase expression level in Tera-1 cells transfected with a vector containing a luciferase reporter gene downstream of the indicated proviral 5’ LTR. LTR activity is expressed as relative light units (RLU; gray) normalized to a control construct with a Renilla luciferase gene driven by an SV40 promoter. The relative promoter activities of the LTR Hs located 551 bp upstream from the 22q11.23 provirus, the 5’ LTR 5B of the 22q11.23 provirus and the 5’ LTR Hs of six other expressed proviruses in Tera-1 cells are shown. (B) Schematic of the 22q11.23 LTR Hs, showing the U3, R and U5 regions. Predicted transcriptional start sites are indicated with black arrows and nucleotide position. Colored boxes indicate previously described promoter element motifs [62,63,64]. Lines below the LTR diagram indicate the regions included in each truncated LTR construct, and numbers to the right of each line indicate the nucleotide position at which the LTR was truncated. GA, GA rich motif (nt 379–386, sequence GGGAAGGG); E, enhancer box (nt 465–476, sequence TTGCAGTTGAGA; nt 485–496, sequence AGGCATCTGTCT; nt 832–843, sequence CTCCATATGCTG); GC, GC rich motif nt 759–763, (sequence CCCCC; nt 602–606, sequence GGCGG); TATA, TATA box (nt 790–797, sequence AATAAATA); Inr, initiator element (nt 807–812, sequence CTCAGA). Cartoon is not drawn to scale. (C) Relative promoter expression levels of truncated 22q11.23 LTR Hs constructs in Tera-1 cells (Kruskal-Wallis, * p < 0.05, ** p < 0.01). All luciferase experiments were conducted in triplicate and are shown as mean ± standard deviation. (D) Schematic of promoter motifs found in the 22q11.21 provirus 5’ LTR Hs, the 22q11.23 LTR Hs and 22q11.23 provirus 5’ LTR 5B. Crossed out boxes indicate presence of a mutation in the motif as compared to the canonical sequence. Cartoon is not drawn to scale.