Fig. 5.

Back mutation of critical transcription factor binding sites to consensus sequences on the 11p15.4 provirus. a, b Multiple sequence alignment of the a ATF and b RORA binding regions on the nine 5′ LTRs of interest in this study as well as a consensus sequence of the site. Sequences are compared against the 11p15.4 5′ LTR site, dots are used for shared identity, and dashes indicate indels. c Relative 5′ LTR promoter activity in HMLE-Ras cells and Hcc1954 cells. Constructs used either contained full ATF and RORA binding sites, or had a binding site removed through back mutation to the consensus sequence. Promoter activity of the 1q22 5′ LTR is shown for comparison. Promoter activity is determined as relative light units (RLU) normalized against the internal control Renilla expression. Statistical significance was generated by ANOVA with Bonferroni’s multiple comparisons test (***p < 0.0005). All experiments were conducted in triplicate and data displayed as the mean ± standard deviation