c-erbA encodes multiple proteins in chicken erythroid cells.
To identify and characterize the proteins encoded by the erbA proto-oncogene, we expressed the C-terminal region of v-erbA in a bacterial trpE expression vector system and used the fusion protein to prepare antiserum. The anti-trp-erbA serum recognized the P75gag-erbA protein encoded by avian erythroblastosis virus and specifically precipitated six highly related proteins ranging in size from 27 to 46 kilodaltons from chicken embryonic erythroid cells. In vitro translation of a chicken erbA cDNA produced essentially the same pattern of proteins. Partial proteolytic maps and antigenicity and kinetic analyses of the in vivo and in vitro proteins indicated that they are related and that the multiple bands are likely to arise from internal initiations within c-erbA to generate a nested set of proteins. All of the c-erbA proteins are predominantly associated with chicken erythroblast nuclei. However, Nonidet P-40 treatment resulted in extraction of the three smaller proteins, whereas the larger proteins were retained. During differentiation of erythroid cells in chicken embryos, we found maximal levels of c-erbA protein synthesis at days 7 to 8 of embryogenesis. By contrast, c-erbA mRNA levels remained essentially constant from days 5 to 12. Together, our results indicate that posttranscriptional or translational mechanisms are involved in regulation of c-erbA expression and in the complexity of its protein products.
Bigler
J
J
Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
Eisenman
R N
RN
D000818:Animals; D015153:Blotting, Western; D002451:Cell Compartmentation; D002467:Cell Nucleus / Q000378:metabolism; D002645:Chickens / Q000235:genetics*; D004912:Erythrocytes / Q000502:physiology*; D004920:Erythropoiesis; D005786:Gene Expression Regulation; D010449:Peptide Mapping; D014176:Protein Biosynthesis; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D011519:Proto-Oncogenes; D012323:RNA Processing, Post-Transcriptional; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014158:Transcription, Genetic
D011518:Proto-Oncogene Proteins; D011993:Recombinant Fusion Proteins