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PMID: 8355708 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The Ets-related transcription factor PU.1 immortalizes erythroblasts.

Molecular and cellular biology ·Vol. 13 ·No. 9 ·1993-09-00 ·Pages 5670-8

Schuetze S, Stenberg PE, Kabat D

Abstract

In vivo studies of Friend virus erythroleukemia have implied that proviral integrations adjacent to the gene for the Ets-related transcription factor PU.1 may inhibit the commitment of erythroblasts to differentiate and cause their capability for indefinite transplantation (C. Spiro, B. Gliniak, and D. Kabat, J. Virol. 62:4129-4135, 1988; R. Paul, S. Schuetze, S. L. Kozak, C. Kozak, and D. Kabat, J. Virol. 65:464-467, 1991). To test this hypothesis, we ligated PU.1 cDNA into a retroviral vector and studied its effects on cultured cells. Infection of fibroblasts with PU.1-encoding retrovirus resulted in PU.1 synthesis followed by nuclear pyknosis, cell rounding, and degeneration. In contrast, in long-term bone marrow cultures, erythroblasts were efficiently and rapidly immortalized. The resulting cell lines were polyclonal populations that contained PU.1, were morphologically blast-like, required erythropoietin and bone marrow stromal cells for survival and proliferation, and spontaneously differentiated at low frequency to synthesize hemoglobin. After 9 months in culture, erythroblasts became stroma independent, and they then grew as clonal cell lines. We conclude that PU.1 perturbs the pathway(s) that controls potential for indefinite proliferation and that it can be used to generate permanent erythroblast cell lines.

Related Genes
EpoR Sfpi-1
MeSH Terms
3T3 Cells Animals Bone Marrow Cells Cell Transformation, Neoplastic Cells, Cultured DNA-Binding Proteins/physiology Erythroblasts/physiology Erythropoiesis Gene Expression Globins/genetics In Vitro Techniques Mice RNA, Messenger/genetics Receptors, Erythropoietin/genetics Retroviridae Proteins, Oncogenic Transfection
Chemicals
DNA-Binding Proteins RNA, Messenger Receptors, Erythropoietin Retroviridae Proteins, Oncogenic v-Spi-1 protein, Friend spleen focus-forming virus Globins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Schuetze S
Department of Biochemistry and Molecular Biology, School of Medicine, Oregon Health Sciences University, Portland 97201-3098.
Stenberg P E
Kabat D
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.13.9.5670-5678.1993
Published
1993-09-00
Pages
5670-8
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC360299
Subset
IM
Grants
NCI NIH HHS · CA25810 · United States
External Links
PubMed source DOI: 10.1128/mcb.13.9.5670-5678.1993 PMC Full Text View journal details
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