Truncated erythropoietin receptor in a murine erythroleukemia cell line.
The Friend spleen focus forming virus produces a 55 kDa envelope glycoprotein which associates with the erythropoietin receptor. We compared the erythropoietin receptor in Friend virus transformed murine erythroleukemic F4N and 707 cell lines with the J2E erythroid line generated by the J2 retrovirus. Reverse transcriptase PCR was used to determine transcript size. Erythropoietin receptor cDNAs were then sequenced and protein products analysed by Western blotting and immunoprecipitation. We show here that the F4N murine erythroleukemic cell line had an enlarged erythropoietin receptor mRNA. In contrast, the 707 and J2E cell line had normal sized transcripts for the receptor. Sequence analysis of the receptor in F4N cells revealed that introns which separate the exons coding for the cytoplasmic domain of the receptor were retained in these transcripts. As a consequence, a premature stop codon had been introduced, leaving only four amino acids in the intracellular portion of the receptor molecule. The normal erythropoietin receptor is approx. 66-70 kDa, but immunoprecipitation of [35S]methionine/cysteine labelled cell lysates with an antibody to the amino-terminus of the erythropoietin receptor identified a truncated 37 kDa protein in F4N cells. Despite the severe carboxy-terminal truncation of the erythropoietin receptor, F4N cells continued to proliferate like the other murine erythroleukemia cell lines. This study shows that failure to remove introns from the erythropoietin receptor mRNA in F4N cells has resulted in the production of a smaller protein with virtually no cytoplasmic domain.
Bittorf
T
T
Department of Biochemistry, University of Western Australia, Nedlands.
Busfield
S J
SJ
Klinken
S P
SP
Tilbrook
P A
PA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral / Q000502:physiology*; D005622:Friend murine leukemia virus / Q000502:physiology*; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism* / Q000821:virology; D051379:Mice; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D017467:Receptors, Erythropoietin / Q000737:chemistry*
D017467:Receptors, Erythropoietin