Apoptosis in hematopoietic cells is associated with an extensive decrease in cellular phosphotyrosine content that can be inhibited by the tyrosine phosphatase antagonist pervanadate.
In the present study, we investigated whether apoptosis in hematopoietic cells is associated with changes in cellular phosphotyrosine content. Murine thymocytes and B cells, human leukemia cells, and normal peripheral blood leukocytes were induced to undergo apoptosis by treatment with specific stimuli or by incubation in growth factor-deprived medium. Multiparameter flow cytometry was used to measure changes in phosphotyrosine content that correlated with the appearance of features of programmed cell death, such as cell shrinkage, DNA fragmentation, and loss of membrane integrity. The results show that conditions that induced apoptosis also induced a dramatic decrease in cellular phosphotyrosine levels. Tyrosine dephosphorylation preceded the loss of plasma membrane integrity and, in most cases, was temporally correlated with the onset of DNA fragmentation. The protein tyrosine phosphatase antagonist pervanadate had a dose-dependent inhibitory effect on both dephosphorylation and apoptosis in murine thymocytes, which were treated with dexamethasone or with the topoisomerase II inhibitor etoposide. The results suggest that extensive tyrosine dephosphorylation is an intrinsic part of the apoptotic process of hematopoietic cells and may be involved mechanistically in the apoptosis induced by certain stimuli.
Becton Dickinson Immunocytometry Systems, San Jose, California, USA. johansen@dnax.org
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F
Lund-Johansen
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T
Frey
J A
JA
Ledbetter
P A
PA
Thompson
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001773:Blood Cells / Q000187:drug effects / Q000502:physiology*; D003907:Dexamethasone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005047:Etoposide / Q000494:pharmacology; D005434:Flow Cytometry; D006410:Hematopoiesis; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D019000:Phosphotyrosine / Q000378:metabolism*; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D013950:Thymus Gland / Q000166:cytology; D014407:Tumor Cells, Cultured; D014638:Vanadates / Q000494:pharmacology*
D004791:Enzyme Inhibitors; C076715:pervanadate; D019000:Phosphotyrosine; D014638:Vanadates; D005047:Etoposide; D003907:Dexamethasone; D017027:Protein Tyrosine Phosphatases