Differential Expression of Low Mr GTP-binding Proteins in Human Megakaryoblastic Leukemia Cell Line, MEG-01, and their Possible Involvement in the Differentiation Process

Koh-ichi Nagata; Yukio Okano; Yoshinori Nozawa

doi:10.1055/s-0038-1655970

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1997; 77(02): 368-375
DOI: 10.1055/s-0038-1655970

Original Article

Schattauer GmbH Stuttgart

Differential Expression of Low Mr GTP-binding Proteins in Human Megakaryoblastic Leukemia Cell Line, MEG-01, and their Possible Involvement in the Differentiation Process

Koh-ichi Nagata

¹The Department of Molecular Pathobiochemistry, Gifu University School of Medicine, Gifu, Japan

,

Yukio Okano

¹The Department of Molecular Pathobiochemistry, Gifu University School of Medicine, Gifu, Japan

,

Yoshinori Nozawa

²The Department of Biochemistry, Gifu University School of Medicine, Gifu, Japan

› Author Affiliations

Abstract

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Summary

The expression of various low Mr GTP-binding proteins at various states of differentiation of a human megakaryoblastic leukemia cell line, MEG-01, was analyzed using thermocycle amplification of mRNA and immunoblotting. MEG-01 cells were found to express mRNAs of rap1A, rap1B, rap2B, ralA, rhoA, rac1, rac2, CDC42Hs, rab1, rab3B, rab6, ram and ran, but not rab4, and the proteins of Rap 1, Rap2, RhoA, Rac1, Rac2, Rab3B, Rab4, Rab6 and Rab8 were expressed. Differentiation of MEG-01 cells induced by 100 nM 12-O-tetradecanoylphorbol-13-acetate revealed the considerable increases in mRNA expression of rap1B, rab3B, rabA, ram and ran whereas the levels of rap2B, rhoA and rac1 decreased. During the differentiation process, significant changes in protein levels of Rap1, RhoA, Rac1, Rac2, Rab3B, Rab4 and Rab6 were observed among three subcellular (cytosol, Triton X-100-soluble membrane and -insoluble cytoskeleton) fractions. The present investigation may be useful for the study of the megakaryocyte differentiation.

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References

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