Figure 5.

The mTERF6 protein is capable of in vitro binding to both DNA and RNA. A, Structural homology modeling of mTERF6. The tertiary structure of human mTERF1 (hMTERF1; deposited as 3MVA) was downloaded from the National Center for Biotechnology Information structure database (http://www.ncbi.nlm.nih.gov/structure) and matched to the predicted structure of mTERF6. The three-dimensional structure of mTERF6 was calculated by I-TASSER (see “Materials and Methods”), and the molecular graphics images were produced using the UCSF Chimera package (see “Materials and Methods”). B, SDS-PAGE showing the purity of recombinant His₆-tagged mTERF6 (mTERF6) expressed in E. coli and purified on nickel-nitrilotriacetic acid agarose. C, Southwestern and northwestern analyses to demonstrate the in vitro binding of His₆-tagged mTERF6 (mTERF6) to dsDNA and RNA, respectively. Shown is the Ponceau Red-stained membrane blotted with increasing concentrations of recombinant mTERF6 protein and a maltose binding protein-tagged control protein. The left and right parts of the membrane were incubated with a radiolabeled total DNA or RNA preparation, respectively.