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. 2020 Jun 8;9:e54995. doi: 10.7554/eLife.54995

Figure 3. Central MARF1 LOTUS domains are required to silence mRNAs containing 3’UTRs of MARF1 target mRNAs.

(A) Schematic diagram of wild-type MARF1 protein, as well C-terminal deletion mutants. Core LOTUS domains required for MARF1-mediated repression are bordered in red. (B–D) RL activity detected in extracts from HEK293 cells expressing the indicated proteins. Cells were cotransfected with constructs expressing the RL-MAML1 reporter or RL-5BoxB reporter, along with FL, and indicated fusion proteins. Histograms represented normalized mean values of RL activity from a minimum of three experiments. RL activity values seen in the presence of λNHA-MARF1ΔNYN were set as 100.

Figure 3—source data 1. Comparative sequence analysis of MARF1 LOTUS domains.
(A) Sequence alignment of conserved amino acids within the MARF1 LOTUS domains 1 through 6 of human (Hs), Drosophila melanogaster (Dm), Xenopus tropicalis (Xt) and Danio rerio (Dr). LOTUS domain amino acids are in bold font. (B) Sequence alignment of human LOTUS domains 3 and 5. Identical amino acids are denoted with an asterisk and conservative amino acids substitutions are denoted with a colon.

Figure 3.

Figure 3—figure supplement 1. Western blot analysis of HEK293 cells expressing λNHA-MARF1 mutants lacking the indicated LOTUS domains.

Figure 3—figure supplement 1.