[Identification and characterization of a novel gene EOLA1 stimulating ECV304 cell proliferation].
OBJECTIVE
To amplify the full-length cDNA and characterize the structure and biological function of a novel expression sequence tag ST55 (GenBank Accession No. BM121646).


METHODS
Rapid amplification of cDNA ends was used to clone the full-length of cDNA of ST55 in this study. Then, its tissue distribution was checked by Northern blots, and the associated protein was screened by GAL 4-based yeast two-hybrid. The effect of stable transfection of the cDNA on cell proliferation was evaluated in ECV304 cells.


RESULTS
A full-length 1404 bp cDNA was cloned, and it was accepted as a novel human mRNA by GenBank (No. AY074889), named endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1). Bioinformatic analysis found that the EOLA1 encoded 158 amino acids, 17.89 kDa protein, and mapped to chromosome Xq27.4 with 5 exons. EOLA1 expressed in different human normal tissues and cancer cell lines. Using the EOLA1 cDNA as bait, we performed a yeast two-hybrid screening of a human liver cDNA library and identified metallothionein 2A (MT2A) as associated protein. The interaction between EOLA1 and MT2A was confirmed by co-immunoprecipitation experiments. Stable transfection of EOLA1 was noted to stimulate ECV304 cell proliferation (P < 0.05).


CONCLUSIONS
The findings suggest that EOLA1 is a novel gene and the interaction of EOLA1 and MT2A may play an important role in cell protection in inflammation reaction.
Liang
Zi-wen
ZW
Department of Endocrinology, Southwest Hospital, Third Military Medical University, Chongqing, PR China. liangzw@mail.tmmu.com.cn
Yang
Zong-cheng
ZC
Liu
Yue-ming
YM
Chen
Yu
Y
Luo
Xiang-dong
XD
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002460:Cell Line; D049109:Cell Proliferation; D041321:Chromosomes, Human, X / Q000235:genetics; D005091:Exons / Q000235:genetics; D006801:Humans; D047468:Immunoprecipitation; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008668:Metallothionein / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D016415:Sequence Alignment; D020798:Two-Hybrid System Techniques
C495142:EOLA1 protein, human; C491053:MT2A protein, human; D008565:Membrane Proteins; D008668:Metallothionein