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. 2018 Aug 9;37(18):e99777. doi: 10.15252/embj.201899777

Figure 1. m5C38‐tRNAAsp GUC dependency by Q in human cell culture.

Figure 1

  1. APB Northern blot using a tRNAHis probe. m5C38 levels of tRNAAsp GUC were measured by 454 bisulfite sequencing at the indicated time points. Both queuosinylation and methylation levels could be restored by the addition of queuine. The slower migration of tRNAHis is eliminated by oxidizing the ribose with periodate, producing a single faster migrating band (ox).
  2. The addition of queuine to HCT116 cells cultured in serum‐free medium resulted in an increase in both queuosinylation and m5C38 levels.
  3. The addition of 20 nM queuine to the SF medium is sufficient to restore tRNAHis QUG in HeLa, whereas 200 nM of q is necessary for queuosinylation of tRNAHis GUG in HCT116 cells.
Data information: S (standard medium), SF (serum‐free medium), q (queuine), Q (queuosine), and G (guanine).Source data are available online for this figure.