Identification of a family of alternatively spliced mRNA species of angiopoietin-1

Citation
Yq. Huang et al., Identification of a family of alternatively spliced mRNA species of angiopoietin-1, BLOOD, 95(6), 2000, pp. 1993-1999
Citations number
29
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Hematology,"Cardiovascular & Hematology Research
Journal title
BLOOD
ISSN journal
0006-4971 → ACNP
Volume
95
Issue
6
Year of publication
2000
Pages
1993 - 1999
Database
ISI
SICI code
0006-4971(20000315)95:6<1993:IOAFOA>2.0.ZU;2-C
Abstract
Angiopoietin-1 (Ang-1) is required for developing vessels, and its absence leads to defects in vessel remodeling. Ang-1 has been identified as the lig and for the tyrosine kinase receptor Tie-a, which is expressed specifically on endothelial cells and early hematopoietic cells. In studying the role o f Tie-2 and Ang-1 in megakaryocytopoiesis, 3 alternatively spliced species of Ang-1 mRNA(Ang-1.3 kb, Ang-0.9 kb, and Ang-0.7 kb) were identified in ad dition to the full-length Ang-1 (Ang-1.5 kb), in the megakaryocyte cell lin e CHRF by reverse transcription-polymerase chain reaction (RT-PCR), and the n cloned and sequenced. The expression of 3 alternatively spliced isoforms of Ang-l was confirmed by RT-PCR using specific primer pairs derived from j unction sites and the 3' end of Ang-1 cDNA, and it was further demonstrated by nuclease protection assay, Northern blotting, and immunoblotting in CHR F cells. Expression of the Ang-1,3 kb isoform was also detected in human pr imary fibroblast cell line FS4, breast cancer cell line MDAMB-463, and CD34 (+)CD41(+) cells of fetal liver and platelets. The function of the 1.5-kb, 1.3-kb, and 0.9-kb isoforms was examined. Recombinant proteins Ang-1.5 and 0.9 kb bind strongly to the recombinant Tie-2 receptor (Tie-2-Fc), whereas the 1,3-kb isoform does not the Ang-1.3 kb isoform binds to the 1.5-kb isof orm, Ang-1.5 kb, but not the 1,3-kb and 0.9-kb isoforms, induces tyrosine p hosphorylation of Tie-5 in human umbilical vein endothelial cells. These da ta suggest that isoforms 1.3 kb and 0.9 kb could serve as dominant negative molecules for the full-length Ang-1, The possible involvement of the newly identified Ang-1 Isoforms in angiogenesis and in growth and differentiatio n of hematopoietic progenitor cells provides a greater complexity to these processes. (C) 2000 by The American Society of Hematology.