Table 1

Growth factor– and eIF4E-dependent polysome recruitment of selected genes

Condition†Cluster 1*
Cluster 3
Cluster 5
Cluster 1: control
Negative control
Igbp1
mEd2
Rnf138
Nap1l1
Cnih
Grwd
Nubp1
Hnrpa1
Uhmk1
Nm23-M2
Ybx1
Fil1
MeanSDMeanSDMeanSDMeanSDMeanSDMeanSDMeanSDMeanSDMeanSDMeanSDMeanSDMeanSD
Percentage of polysome association                         
    Empty vector                         
        ss 50.6 6.4 30.2 6.3 26.6 9.2 43.3 7.8 16.0 3.8 79.3 8.2 31.8 1.2 41.0 9.6 47.4 1.6 47.5 3.4 34.1 3.9 85.3 2.0 
        NF 4.1 1.5 2.7 0.6 3.1 2.1 2.6 1.4 <2 — 45.5 2.8 5.8 1.6 6.7 1.4 10.2 2.4 3.1 0.9 3.8 0.5 43.0 11.8 
        E 18.0 1.3 8.7 2.2 37.3 7.0 14.1 6.0 7.5 2.8 44.5 8.5 29.9 7.9 16.3 0.2 14.5 1.1 38.9 3.4 18.0 0.8 70.7 5.3 
        S 26.1 5.7 15.1 2.4 40.6 12.5 35.3 6.2 8.1 0.2 76.5 8.0 28.9 4.0 33.1 3.1 36.5 2.9 50.0 6.3 17.2 2.1 64.4 3.8 
        ES 64.8 12.3 26.7 7.8 54.1 22.9 49.7 23.3 19.2 4.9 75.3 4.9 44.8 11.4 61.2 7.6 38.4 1.1 79.3 6.2 48.8 3.1 72.9 9.1 
        S + LY <2 — 3.1 0.6 <2 — 10.8 5.9 <2 — 49.1 5.7 <2 — 2.3 2.7 11.7 3.0 2.0 0.4 2.1 0.3 36.3 11.7 
        S + rap 5.9 2.0 7.4 2.6 34.6 22.9 24.4 18.3 <2 — 59.0 1.9 12.0 3.9 13.6 0.4 18.9 5.8 16.8 2.7 8.8 1.3 50.1 6.0 
        E + LY <2 — <2 — <2 — 3.8 4.5 <2 — 22.9 3.0 26.1 0.5 3.5 0.2 4.6 1.6 8.4 1.4 4.4 0.9 42.3 5.6 
        E + rap 3.6 1.2 7.8 3.4 5.6 3.4 16.2 5.5 7.9 2.7 65.5 2.3 18.2 7.8 16.4 0.6 15.7 4.9 18.1 0.6 14.2 3.3 65.6 2.4 
    eIF4E overexpression                         
        NF 22.8 1.3 16.1 3.8 28.8 7.3 22.7 5.9 25.9 9.0 64.1 11.6 17.6 2.3 30.3 4.3 72.1 1.1 20.8 2.2 16.9 1.5 75.7 3.5 
        S 36.5 2.9 23.9 4.2 50.9 2.1 55.0 12.6 47.4 7.5 92.2 14.8 63.5 0.5 59.7 8.0 60.7 3.5 53.5 15.3 21.8 2.2 85.1 5.2 
        S + LY 28.0 13.6 10.1 4.9 12.6 6.3 12.5 0.2 17.4 14.8 79.4 5.6 38.1 9.4 32.2 5.7 61.0 1.7 20.3 8.1 9.5 1.7 65.1 8.8 
Relative gene expression (fold change)                         
    Total RNA normalized to NF                         
        E 1.6 0.5 2.4 0.1 2.8 0.3 1.1 0.0 1.2 0.4 7.0 1.7 2.6 0.2 0.5 0.2 1.4 0.1 1.8 0.3 1.3 0.2 0.7 0.1 
        S 2.1 0.5 2.3 0.1 2.2 0.3 1.3 0.1 1.3 0.3 5.8 0.1 2.5 0.3 0.6 0.0 1.2 0.1 0.7 0.1 0.9 0.1 0.8 0.1 
        ES 2.2 0.6 2.3 0.2 2.5 0.0 1.8 0.2 1.9 0.1 7.2 0.3 3.2 0.4 0.5 0.0 0.9 0.1 1.3 0.1 1.5 0.2 0.8 0.1 
    pb RNA normalized to NF                         
        E 4.7 0.4 4.0 0.2 3.0 0.4 7.0 2.1 5.0 1.0 1.0 0.0 3.6 0.9 2.6 0.5 1.6 0.3 8.0 1.1 5.0 0.5 1.5 0.1 
        S 5.7 0.1 5.8 0.5 9.0 0.9 12.0 5.1 6.0 1.7 1.7 0.1 6.4 1.4 5.1 0.6 3.0 0.2 11.5 0.7 5.0 0.2 1.2 0.0 
        ES 14.9 2.6 13.5 1.5 21.5 1.1 9.2 2.5 17.3 2.3 1.7 0.0 16.2 2.7 9.7 2.4 3.2 0.0 14.0 3.2 11.7 0.3 1.3 0.1 
    Total RNA normalized to ss                         
        diff t24 1.2 0.1 1.1 0.1 3.4 0.1 0.5 0.1 1.3 0.02 1.7 0.2 0.6 0.3 1.1 0.1 1.3 0.1 0.8 0.1 0.8 0.2 1.2 0.0 
        diff t48 0.5 0.0 1.1 0.0 1.7 0.1 0.5 0.1 0.8 0.1 0.8 0.1 0.4 0.2 0.9 0.1 1.3 0.2 0.5 0.0 0.5 0.1 1.1 0.1 
        diff t72 0.7 0.0 0.4 0.0 0.6 0.0 0.3 0.2 0.7 0.0 0.3 0.0 0.4 0.0 0.5 0.1 0.4 0.1 0.4 0.1 0.3 0.1 1.4 0.1 
Condition†Cluster 1*
Cluster 3
Cluster 5
Cluster 1: control
Negative control
Igbp1
mEd2
Rnf138
Nap1l1
Cnih
Grwd
Nubp1
Hnrpa1
Uhmk1
Nm23-M2
Ybx1
Fil1
MeanSDMeanSDMeanSDMeanSDMeanSDMeanSDMeanSDMeanSDMeanSDMeanSDMeanSDMeanSD
Percentage of polysome association                         
    Empty vector                         
        ss 50.6 6.4 30.2 6.3 26.6 9.2 43.3 7.8 16.0 3.8 79.3 8.2 31.8 1.2 41.0 9.6 47.4 1.6 47.5 3.4 34.1 3.9 85.3 2.0 
        NF 4.1 1.5 2.7 0.6 3.1 2.1 2.6 1.4 <2 — 45.5 2.8 5.8 1.6 6.7 1.4 10.2 2.4 3.1 0.9 3.8 0.5 43.0 11.8 
        E 18.0 1.3 8.7 2.2 37.3 7.0 14.1 6.0 7.5 2.8 44.5 8.5 29.9 7.9 16.3 0.2 14.5 1.1 38.9 3.4 18.0 0.8 70.7 5.3 
        S 26.1 5.7 15.1 2.4 40.6 12.5 35.3 6.2 8.1 0.2 76.5 8.0 28.9 4.0 33.1 3.1 36.5 2.9 50.0 6.3 17.2 2.1 64.4 3.8 
        ES 64.8 12.3 26.7 7.8 54.1 22.9 49.7 23.3 19.2 4.9 75.3 4.9 44.8 11.4 61.2 7.6 38.4 1.1 79.3 6.2 48.8 3.1 72.9 9.1 
        S + LY <2 — 3.1 0.6 <2 — 10.8 5.9 <2 — 49.1 5.7 <2 — 2.3 2.7 11.7 3.0 2.0 0.4 2.1 0.3 36.3 11.7 
        S + rap 5.9 2.0 7.4 2.6 34.6 22.9 24.4 18.3 <2 — 59.0 1.9 12.0 3.9 13.6 0.4 18.9 5.8 16.8 2.7 8.8 1.3 50.1 6.0 
        E + LY <2 — <2 — <2 — 3.8 4.5 <2 — 22.9 3.0 26.1 0.5 3.5 0.2 4.6 1.6 8.4 1.4 4.4 0.9 42.3 5.6 
        E + rap 3.6 1.2 7.8 3.4 5.6 3.4 16.2 5.5 7.9 2.7 65.5 2.3 18.2 7.8 16.4 0.6 15.7 4.9 18.1 0.6 14.2 3.3 65.6 2.4 
    eIF4E overexpression                         
        NF 22.8 1.3 16.1 3.8 28.8 7.3 22.7 5.9 25.9 9.0 64.1 11.6 17.6 2.3 30.3 4.3 72.1 1.1 20.8 2.2 16.9 1.5 75.7 3.5 
        S 36.5 2.9 23.9 4.2 50.9 2.1 55.0 12.6 47.4 7.5 92.2 14.8 63.5 0.5 59.7 8.0 60.7 3.5 53.5 15.3 21.8 2.2 85.1 5.2 
        S + LY 28.0 13.6 10.1 4.9 12.6 6.3 12.5 0.2 17.4 14.8 79.4 5.6 38.1 9.4 32.2 5.7 61.0 1.7 20.3 8.1 9.5 1.7 65.1 8.8 
Relative gene expression (fold change)                         
    Total RNA normalized to NF                         
        E 1.6 0.5 2.4 0.1 2.8 0.3 1.1 0.0 1.2 0.4 7.0 1.7 2.6 0.2 0.5 0.2 1.4 0.1 1.8 0.3 1.3 0.2 0.7 0.1 
        S 2.1 0.5 2.3 0.1 2.2 0.3 1.3 0.1 1.3 0.3 5.8 0.1 2.5 0.3 0.6 0.0 1.2 0.1 0.7 0.1 0.9 0.1 0.8 0.1 
        ES 2.2 0.6 2.3 0.2 2.5 0.0 1.8 0.2 1.9 0.1 7.2 0.3 3.2 0.4 0.5 0.0 0.9 0.1 1.3 0.1 1.5 0.2 0.8 0.1 
    pb RNA normalized to NF                         
        E 4.7 0.4 4.0 0.2 3.0 0.4 7.0 2.1 5.0 1.0 1.0 0.0 3.6 0.9 2.6 0.5 1.6 0.3 8.0 1.1 5.0 0.5 1.5 0.1 
        S 5.7 0.1 5.8 0.5 9.0 0.9 12.0 5.1 6.0 1.7 1.7 0.1 6.4 1.4 5.1 0.6 3.0 0.2 11.5 0.7 5.0 0.2 1.2 0.0 
        ES 14.9 2.6 13.5 1.5 21.5 1.1 9.2 2.5 17.3 2.3 1.7 0.0 16.2 2.7 9.7 2.4 3.2 0.0 14.0 3.2 11.7 0.3 1.3 0.1 
    Total RNA normalized to ss                         
        diff t24 1.2 0.1 1.1 0.1 3.4 0.1 0.5 0.1 1.3 0.02 1.7 0.2 0.6 0.3 1.1 0.1 1.3 0.1 0.8 0.1 0.8 0.2 1.2 0.0 
        diff t48 0.5 0.0 1.1 0.0 1.7 0.1 0.5 0.1 0.8 0.1 0.8 0.1 0.4 0.2 0.9 0.1 1.3 0.2 0.5 0.0 0.5 0.1 1.1 0.1 
        diff t72 0.7 0.0 0.4 0.0 0.6 0.0 0.3 0.2 0.7 0.0 0.3 0.0 0.4 0.0 0.5 0.1 0.4 0.1 0.4 0.1 0.3 0.1 1.4 0.1 

Mean values are from at least 3 measurements.

E indicates erythropoietin; SCF, stem cell factor; ES, E + SCF; NF, no factor; LY, LY294002; ss, steady-state proliferation conditions; rap, rapamycin; diff t, diffrentiation time (hours); and —, not applicable.

*

Selected genes correspond to cluster 1, 3, or 5 in Figure 1. Nm23-M2 and Ybx1 are present in cluster 1 but were already known to be controlled by PI3K and eIF4E; Fli-1 represents global effects on polysome recruitment.

†

Erythroid progenitors were grown under steady-state proliferation conditions (ss), factor-deprived (4 hours), and restimulated for 2 hours in the absence of factor (NF), in the presence of Epo (E, 5 U/mL), SCF (S, 500 ng/mL), or both factors (ES). In some instances, cells were treated with LY294002 (LY, 15 mm) or rapamycin (rap, 50 ng/mL) during stimulation. Alternatively, cells were induced to differentiate for 24, 48, or 72 hours.

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