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
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
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.