FIG. 7.
LANA and MeCP2 cooperate to regulate transcription. (A) LANA bound to reiterated copies of the KSHV TR sequence can repress a constitutively active promoter located on the same plasmid. The schematic shows MeCP2 truncations, indicating the three domains (MBD, CRID, and TRD) required for association with LANA. (B) HeLa cells were cotransfected with the pGL3p-7xTR reporter and expression plasmids encoding derivatives of LANA and/or MeCP2. Luciferase activity was measured after 24 h, and the degree of repression (n-fold) was calculated relative to expression in cells with the reporter alone. Values represent the means and standard deviations of results from three independent assays. The dotted line indicates the level of repression by LANANC alone. Note that expression of MeCP2 in the absence of LANA results in modest stimulation. (C) Results of a repression assay comparing the LANAFL wild type (WT) and the CBM deletion mutant (Δ22) in the presence or absence of MeCP2. (D) Stimulation of the human E2F1 promoter by LANA and MeCP2 is sensitive to mutations in the LANA CBM. Cells were cotransfected with the pE2F1-luc reporter and plasmids expressing WT and mutant versions of LANANC in the presence or absence of MeCP2FL. The dotted line corresponds to reporter activity in the presence of LANANC alone.