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. Author manuscript; available in PMC: 2012 Dec 27.
Published in final edited form as: Nature. 2012 Jul 12;487(7406):244–248. doi: 10.1038/nature11126

Figure 3. Biosynthesis of hypusine as a tumour suppressor pathway.

Figure 3

a, Schematic of the polyamine–hypusine pathway. Enzymes and compounds are indicated as follows: ODC1, ornithine decarboxylase; SRM, spermidine synthase; SMS, spermine synthase; DHPS, deoxyhypusine synthase; DOHH, deoxyhypusine hydrolase; SMO(X), spermine oxidase; PAOX, polyamine oxidase; SSAT, spermidine–spermine acetyltransferase; DAX, diamine transporter. SAM, S-adenosylmethionine; dc-SAM, decarboxylated S-adenosylmethionine. The SSAT–PAOX axis can also convert spermidine to putrescine (not shown). b, Survival curves for mice (n = 10 per pool) transduced with the following shRNAs or shRNA pools: Srm (violet, P < 0.001), Dhps (blue, P < 0.001), Amd1 (Amd1.2606, green, P < 0.001) and control (luc.1309, black). c, Two-dimensional PAGE followed by eIF5A immunoblotting of lymphomas driven by the indicated shRNAs. The p53.1224 lymphomas were treated with 10 µM N1-guanyl-1,7-diaminoheptane (GC-7) in short-term culture conditions. Arrows indicate the hypusinated form. d, Quantification of hypusinated/total eIF5A ratio for the indicated shRNAs (n = 3 per group, *P < 0.05). Error bars, s.d. e, Lymphomas generated by transduction of Eif5a shRNAs were retrovirally transduced with control vector (black), a complementary DNA encoding wild-type Eif5a (red) or a mutant complementary DNA that cannot be hypusinated because of the substitution of the modified lysine (K50A, blue). Cherry percentages were monitored for 5 days and normalized to the Cherry fraction at day 1 (100). Error bars, s.d. (n = 4 for each time point, *P < 0.05). f, Annexin V+/PI− fractions of GFP-positive B cells 3 weeks after adoptive transplant of Eμ-myc HSPCs transduced with the indicated shRNAs or shRNA pools (n = 3 per each shRNA; **P < 0.01). Error bars, s.d. g, Western blot analysis for expression of the indicated proteins in Eμ-myc pre-malignant B cells transduced with the indicated shRNAs and sorted 21 days after transplant. Cells from three mice were pooled for each shRNA.