Isolation of Mouse Ang-vl Gene, Overexpressed in the NIH3T3 Transformed by the PAX3/FKHR Chimeric Transcription Factor

Jeon, In Sang; Shapiro, David N

Abstract

BACKGROUND: Angiogenin, a 14.1-kDa protein isolated from the medium conditioned by HT-29 human colon carcinoma cells, induces the angiogenesis. In contrast to the human angiogenin, thought to have one homologue, the mouse angiogenin is known to have several angiogenin homologues. During we were investigating the target genes, overexpressed by the chimeric PAX3/FKHR transcription factor, new gene closely similar to the mouse angiogenin rather than angiogenin itself was obtained. We report this Ang-vl gene to understand the process of angiogenesis by comparing the mouse angiogenin family genes.
METHODS
The representational difference analysis was used to investigate the target genes over expressed by the PAX3/FKHR chimeric transcription factor. The target genes were subcloned into the pBluescriptSK + and sequenced using the 73 and 77 vector itself primers. Analyses of the completed consensus nucleic acid and peptide sequences were performed using the intelligenetics and GCG software packages as well as BLAST algorithms.
RESULTS
The Ang-vl gene, including the glutamine that becomes the N-terminal amino acid by the post-translational peptidase reaction and stop codon, was obtained.
CONCLUSIONS
We cloned the one member of the mouse angiogenin family genes. From the point of protein chemistry, the mechanism of angiogenin or, for that matter, of any other blood vessel inducing proteins is not yet known. However, the homologues of the angiogenin might interact each other to regulate the angiogenesls. In this regard, the Ang-vl gene provides an opportunity to understand the mechanism of angiogenesis.