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Differential regulation of extracellular signal-regulated kinase 1 and 2 activity during 12-O-tetradecanoylphorbol 13-acetate-induced differentiation of HL-60 cells.

Abstract：

:In this study we have analyzed short- and long-term changes in extracellular signal-regulated kinase (ERK) 1 and 2 activity during 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced differentiation of human promyelocytic leukemia cells. Immunoprecipitation of HL-60 cellular extracts with an ERK antibody followed by in vitro myelin basic protein phosphorylation demonstrated a rapid reduction in total ERK activity by 70%. Mitogen-activated protein kinase substrate peptide phosphorylation also demonstrated that this reduction was sustained during differentiation. Immunoblot analysis revealed that ERK1 and ERK2 are the predominant ERK isoforms present in HL-60 cells and that over a 96-h period ERK1 protein was gradually reduced by 60% while ERK2 protein showed only a small, insignificant reduction. Therefore, the large, rapid decrease in total ERK activity could not be attributed to the gradual reductions in ERK1 or ERK2 amounts. Immunoblot analysis with two different phosphotyrosine antibodies revealed a rapid decrease in ERK1 phosphotyrosine and a concurrent transient increase in ERK2 phosphotyrosine. These contrasting changes in phosphorylated ERKs were paralleled by respective shifts in mobility during SDS-PAGE analysis. Together these results indicate that the rapid reduction in total ERK activity is due to rapid tyrosine and possible threonine dephosphorylation of ERK1 but not of ERK2. These results also indicate that ERK1 and ERK2 are regulated by distinct mechanisms during TPA-induced HL-60 differentiation, suggesting that their biological roles are nonredundant.

journal_name

Exp Cell Res

journal_title

Experimental cell research

authors

Meighan-Mantha RL,Wellstein A,Riegel AT

doi

10.1006/excr.1997.3631

subject

Has Abstract

pub_date

1997-08-01 00:00:00

pages

321-8

issue

2

eissn

0014-4827

issn

1090-2422

pii

S0014-4827(97)93631-4

journal_volume

234

pub_type

杂志文章

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