Cloning and structural characterization of ECTACC, a new member of the transforming acidic coiled coil (TACC) gene family: cDNA sequence and expression analysis in human microvascular endothelial cells

Citation
Jj. Pu et al., Cloning and structural characterization of ECTACC, a new member of the transforming acidic coiled coil (TACC) gene family: cDNA sequence and expression analysis in human microvascular endothelial cells, CYTOKINE, 13(3), 2001, pp. 129-137
Citations number
37
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Cell & Developmental Biology
Journal title
CYTOKINE
ISSN journal
1043-4666 → ACNP
Volume
13
Issue
3
Year of publication
2001
Pages
129 - 137
Database
ISI
SICI code
1043-4666(20010207)13:3<129:CASCOE>2.0.ZU;2-2
Abstract
Erythropoietin (Epo) transduces mitogenic and chemoattractant signals to hu man endothelial cells, Identifications of Epo-responsive genes are importan t for understanding the molecular nature of Epo signaling in endothelial ce lls. The effects of Epo on differential expression of various genes were ex amined in human microvascular endothelial cells (HMVEC) by differential dis play reverse transcriptase polymerase chain reaction (RT-PCR). In the curre nt study we obtained from Epo-treated HMVEC a cDNA fragment with characteri stics of the 3' end of mRNA. Using the cDNA fragment, we then selectively i solated a full-length clone by screening an unamplified endothelial cell cD NA library followed by 5' rapid amplification of cDNA ends by polymerase ch ain reaction (RACE-PCR). The nucleotide sequence of the longest cDNA reveal ed an open reading frame of 3311 nucleotides that encodes a protein consist ing of similar to 906 amino acids with a predicted MW of similar to 100 kDa . The nucleotide sequence of the cDNA is nearly identical to that of transf orming acidic coiled coil-containing (TACC2) and anti-zuai-1 (AZU-1) cDNA c lones except at the 5'- and 3'-ends. Northern blot analysis showed an incre ase in endothelial-TACC-related mRNA levels in Epo-treated cells in compari son to that of the control cells. Endothelial-TACC-related mRNA was highly expressed in heart and skeletal muscle tissue. Placenta and brain tissue ex hibited low levels of expression of endothelial-TACC-related gene. Southern blot analysis of genomic DNA from somatic cell hybrids showed that endothe lial-TACC-related cDNA maps to chromosome 10. Immunofluorescence microscopy and the occurrence of several putative phosphorylation and SH3 binding sit es on the deduced protein suggest that endothelial-TACC-related protein may be involved in Epo signaling cascades in endothelial cells. (C) 2001 Acade mic Press.