Average inter-ribosome distance on mRNA in U2OS and HEK293 cells

Range ~200 - 400 nts
Organism Human Homo sapiens
Reference Yan X, Hoek TA, Vale RD, Tanenbaum ME. Dynamics of Translation of Single mRNA Molecules In Vivo. Cell. 2016 May 5 165(4):976-89. doi: 10.1016/j.cell.2016.04.034. p.979 right column top paragraphPubMed ID27153498
Method P.987 left column 4th paragraph: "In summary, here [investigators] have developed an imaging method that enables the measurement of ribosome initiation and translocation rates on single mRNA molecules in live cells." P.987 left column bottom paragraph: "U2OS [human osteosarcoma U2OS cell line] and HEK293 [Human Embryonic Kidney 293 cell] cells were grown in DMEM/5% with Pen/Strep."
Comments P.979 right column top paragraph: "The translation site scFv-GFP [GFP-tagged single-chain intracellular antibody] signal did not substantially increase after CHX treatment (Figure S2C), indicating that under [investigators'] experimental conditions, the lag time between peptide synthesis and scFv-GFP binding does not detectably affect translation-site intensity. Based on the above controls and [their] mathematical model, [they] could estimate the ribosome number per mRNA from the fluorescence intensity of the translation site. Approximately 30% of the mRNAs did not have a corresponding GFP signal, suggesting that they were not actively translating. For the remaining 70% of the mRNAs that were translating, the majority (76%) had between 10–25 ribosomes (Figure 2A Supplemental Experimental Procedures), corresponding to an average inter-ribosome distance of ∼200–400 nucleotides (nt)."
Entered by Uri M
ID 112743