TY - JOUR
T1 - V-erbA generates ribosomes devoid of RPL11 and regulates translational activity in avian erythroid progenitors
AU - Nguyen-Lefebvre, A. T.
AU - Leprun, G.
AU - Morin, V.
AU - Viñuelas, J.
AU - Couté, Y.
AU - Madjar, J. J.
AU - Gandrillon, O.
AU - Gonin-Giraud, S.
N1 - Funding Information:
We thank Dr Stephanie Gobert-Gosse (Universite Claude Bernard Lyon 1, Villeurbanne, France) for her expert advice on the two-dimensional electrophoresis experiments and Clément Soleilhavoup for his technical help in the analysis of total mRNA levels by RT–qPCR. This work received support from grants from Association pour la Recherche contre le Cancer, Ligue contre le Cancer (Comité Départemental du Rhône), Société Franc¸aise d’Hématologie (SFH), Région Rhône-Alpes, Université Claude Bernard Lyon 1 (UCBL1) and Centre National de la Recherche Scientifique (CNRS).
PY - 2014/3/20
Y1 - 2014/3/20
N2 - The v-erbA oncogene transforms chicken erythrocytic progenitors (T2EC) by blocking their differentiation and freezing them in a state of self-renewal. Transcriptomes of T2EC, expressing either v-erbA or a non-transforming form of v-erbA (S61G), were compared using serial analysis of gene expression and some, but not all, mRNA-encoding ribosomal proteins were seen to be affected by v-erbA. These results suggest that this oncogene could modulate the composition of ribosomes. In the present study, we demonstrate, using two-dimensional difference in gel electrophoresis, that v-erbA-expressing cells have a lower amount of RPL11 associated with the ribosomes. The presence of ribosomes devoid of RPL11 in v-erbA-expressing cells was further confirmed by immunoprecipitation. In order to assess the possible impact of these specialized ribosomes on the translational activity, we analyzed proteomes of either v-erbA or S61G-expressing cells using 2D/mass spectrometry, and identified nine proteins present in differing amounts within these cells. Among these proteins, we focused on HSP70 because of its involvement in erythroid differentiation. Our results indicate that, in v-erbA-expressing cells, hsp70 is not only transcribed but also translated more efficiently, as shown by polyribosome fractionation experiments. We demonstrate here, for the first time, the existence of ribosomes with different protein components, notably ribosomes devoid of RPL11, and a regulation of mRNA translation depending on v-erbA oncogene expression.
AB - The v-erbA oncogene transforms chicken erythrocytic progenitors (T2EC) by blocking their differentiation and freezing them in a state of self-renewal. Transcriptomes of T2EC, expressing either v-erbA or a non-transforming form of v-erbA (S61G), were compared using serial analysis of gene expression and some, but not all, mRNA-encoding ribosomal proteins were seen to be affected by v-erbA. These results suggest that this oncogene could modulate the composition of ribosomes. In the present study, we demonstrate, using two-dimensional difference in gel electrophoresis, that v-erbA-expressing cells have a lower amount of RPL11 associated with the ribosomes. The presence of ribosomes devoid of RPL11 in v-erbA-expressing cells was further confirmed by immunoprecipitation. In order to assess the possible impact of these specialized ribosomes on the translational activity, we analyzed proteomes of either v-erbA or S61G-expressing cells using 2D/mass spectrometry, and identified nine proteins present in differing amounts within these cells. Among these proteins, we focused on HSP70 because of its involvement in erythroid differentiation. Our results indicate that, in v-erbA-expressing cells, hsp70 is not only transcribed but also translated more efficiently, as shown by polyribosome fractionation experiments. We demonstrate here, for the first time, the existence of ribosomes with different protein components, notably ribosomes devoid of RPL11, and a regulation of mRNA translation depending on v-erbA oncogene expression.
KW - 2D-DIGE
KW - Proteomic
KW - Ribosomal proteins
KW - Translational activity regulation
KW - V-erbA oncogene
UR - http://www.scopus.com/inward/record.url?scp=84897061004&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84897061004&partnerID=8YFLogxK
U2 - 10.1038/onc.2013.93
DO - 10.1038/onc.2013.93
M3 - Article
C2 - 23563180
AN - SCOPUS:84897061004
SN - 0950-9232
VL - 33
SP - 1581
EP - 1589
JO - Oncogene
JF - Oncogene
IS - 12
ER -