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Blood, 15 August 2000, Vol. 96, No. 4, pp. 1366-1373
HEMATOPOIESIS
Hematopoietic-specific  1 tubulin participates in a pathway of
platelet biogenesis dependent on the transcription factor
NF-E2
Patrick Lecine,
Joseph E. Italiano Jr,
Sang-We Kim,
Jean-Luc Villeval, and
Ramesh A. Shivdasani
From the Departments of Adult Oncology and Cancer
Biology, Dana-Farber Cancer Institute, and the Department of Medicine,
Brigham & Women's Hospital and Harvard Medical School, Boston, MA.
The cellular and molecular bases of platelet release by terminally
differentiated megakaryocytes represent important questions in cell
biology and hematopoiesis. Mice lacking the transcription factor NF-E2
show profound thrombocytopenia, and their megakaryocytes fail to
produce proplatelets, the microtubule-based precursors of blood
platelets. Using mRNA subtraction between normal and NF-E2-deficient
megakaryocytes, cDNA was isolated encoding  1 tubulin, the most
divergent tubulin isoform. In NF-E2-deficient megakaryocytes, 1
tubulin mRNA and protein are virtually absent. The expression of 1
tubulin is exquisitely restricted to platelets and megakaryocytes,
where it appears late in differentiation and localizes to microtubule
shafts and coils within proplatelets. Restoring NF-E2 activity in a
megakaryoblastic cell line or in NF-E2-deficient primary
megakaryocytes rescues the expression of 1 tubulin. Re-expressing
1 tubulin in isolation does not, however, restore proplatelet
formation in the defective megakaryocytes, indicating that other
critical factors are required; indeed, other genes identified by mRNA
subtraction also encode structural and regulatory components of the
cytoskeleton. These findings provide critical mechanistic links between
NF-E2, platelet formation, and selected microtubule proteins, and they
also provide novel molecular insights into thrombopoiesis.
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