(A) Three capped mRNAs were used to test TIE-mediated inhibition in vitro. Hoxa3 TIE and Hoxa11 TIE were placed upstream of the 5’UTR of Hbb-b1 and the Renilla luciferase coding sequence. …
(A) Translation efficiency was measured with a 5’UTR Hbb-b1 Renilla luciferase (RLuc) reporter without TIE. A-capped and m7G-capped mRNAs were first used to assess cap dependency of the rabbit …
The structures of (A) Hoxa3 TIE (170 nucleotides) and (B) Hoxa11 TIE (216 nucleotides) were obtained by chemical probing using base-specific reagents, dimethyl sulfate (DMS) and …
Probing experiments by DMS and CMCT were performed in triplicates. Figures S2A and S2B represent averages of reactivities of DMS and CMCT for Hoxa3 TIE, respectively. Figures S2C and S2D represent …
(A) Substitution mutations in uAUG111and uAUG123 to UAC in Hoxa3 TIE were performed. Constructs with the corresponding mutations were translated in rabbit reticulocyte lysate (RRL) and luciferase …
(A) (To the left) AUG-like mutations (AUU, GUG, CUG, and ACG) were tested in vitro in rabbit reticulocyte lysate (RRL) compared to wt Hoxa3. (To the right) Representation of uAUG111 context in the …
(A) Three transcripts were used for this experiment: full-length 5’UTR of Hoxa3, a deletion mutant at nucleotide G333 in Hoxa3 internal ribosome entry site (IRES) and a control transcript without …
(A) A single nucleotide deletion of A220 in Hoxa3 TIE is performed to frameshift the uORF frame to the same as the main Renilla luciferase (RLuc) ORF. Another mutation was performed combining this …
(A) Mutational analysis of uAUGs in Hoxa11 TIE. Three transcripts with AUG/UAC mutations were used: M1: (AUG/UAC)84 + (AUG/UAC)159, M2: (AUG/UAC)84, and M3: AUG/UAC159. Transcripts were translated …
The stem loop structure of Hoxa11 TIE (104–154) was transplanted in the 5’UTR of Hbb-b1 with 25 nucleotides spanning from each extremity. The three shown transcripts were in vitro translated in …
Polysome fractionation of pre-initiation complexes programmed with m7G-capped radioactive mRNAs containing Wt Hoxa3 translation inhibitory element (TIE) and the mutant (AUG/UAC)84 in rabbit …
Mass spectrometry analysis of cycloheximide-blocked translation initiation complexes and on three transcripts: Hoxa3 TIE, Hoxa11 TIE placed upstream of 5’UTR of Hbb-b1, and the 5’UTR of Hbb-b1 …
Mass spectrometry analysis of GMP-PNP-blocked translation initiation complexes and on three transcripts: Hoxa3 TIE, Hoxa11 TIE placed upstream of 5’UTR of Hbb-b1, and the 5’UTR of Hbb-b1 (control). …
Mutational analysis of A-motif sequences in Hoxa3 TIE shows a requirement for an upstream A-motif for efficient inhibition. (A) Co-transfection assay was performed using pmirGLO plasmids with …
A model for Hoxa3 TIE suggests a ribosomal assembly on the uAUG111 with a requirement of eIF2D initiation factor. The model for Hoxa11 TIE suggests a stalled 80S ribosome on AUG-stop codons …
(A) Nucleotides 1–170 from the mouse Hoxa3 TIE sequence were aligned with different species. The position of uAUG in Mus musculus is highlighted in a red box. Accession numbers: Felis catus: …
Mass spectrometry analysis of pre-initiation complexes programmed with Hoxa3 TIE, Hoxa11 TIE, and 5’UTR Hbb-b1 in the presence of cycloheximide.
Mass spectrometry analysis of pre-initiation complexes programmed with Hoxa3 TIE, Hoxa11 TIE, and 5’UTR Hbb-b1 in the presence of GMP-PNP.