Transferrin receptor hyperexpression in primary erythroblasts is lost on transformation by avian erythroblastosis virus.
In primary chicken erythroblasts (stem cell factor [SCF] erythroblasts), transferrin receptor (TfR) messenger RNA (mRNA) and protein were hyperexpressed as compared to nonerythroid chicken cell types. This erythroid-specific hyperexpression was abolished in transformed erythroblasts (HD3E22 cells) expressing the v-ErbA and v-ErbB oncogenes of avian erythroblastosis virus. TfR expression in HD3E22 cells could be modulated by changes in exogenous iron supply, whereas expression in SCF erythroblasts was not subject to iron regulation. Measurements of TfR mRNA half-life indicated that hyperexpression in SCF erythroblasts was due to a massive stabilization of transcripts even in the presence of high iron levels. Changes in mRNA binding activity of iron regulatory protein 1 (IRP1), the primary regulator of TfR mRNA stability in these cells, correlated well with TfR mRNA expression; IRP1 activity in HD3E22 cells and other nonerythroid cell types tested was iron dependent, whereas IRP1 activity in primary SCF erythroblasts could not be modulated by iron administration. Analysis of avian erythroblasts expressing v-ErbA alone indicated that v-ErbA was responsible for these transformation-specific alterations in the regulation of iron metabolism. In SCF erythroblasts high amounts of TfR were detected on the plasma membrane, but a large fraction was also located in early and late endosomal compartments, potentially concealing temporary iron stores from the IRP regulatory system. In contrast, TfR was almost exclusively located to the plasma membrane in HD3E22 cells. In summary, stabilization of TfR mRNA and redistribution of Fe-Tf/TfR complexes to late endosomal compartments may contribute to TfR hyperexpression in primary erythroblasts, effects that are lost on leukemic transformation.
Lobmayr
Lioba
L
Institute of Medical Biochemistry, Division of Molecular Biology, Vienna Biocenter, Vienna, Austria.
Sauer
Thomas
T
Killisch
Iris
I
Schranzhofer
Matthias
M
Wilson
Robert B
RB
Ponka
Prem
P
Beug
Hartmut
H
Müllner
Ernst W
EW
D001355:Alpharetrovirus; D000818:Animals; D002451:Cell Compartmentation; D002472:Cell Transformation, Viral / Q000502:physiology*; D002645:Chickens; D004900:Erythroblasts / Q000378:metabolism* / Q000821:virology*; D006706:Homeostasis; D007501:Iron / Q000378:metabolism / Q000493:pharmacokinetics / Q000494:pharmacology; D035941:Iron Regulatory Protein 1; D035925:Iron-Regulatory Proteins; D007506:Iron-Sulfur Proteins / Q000378:metabolism; D021381:Protein Transport; D020871:RNA Stability; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D011990:Receptors, Transferrin / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*
D035925:Iron-Regulatory Proteins; D007506:Iron-Sulfur Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011990:Receptors, Transferrin; D007501:Iron; D035941:Iron Regulatory Protein 1