Identification and characterization of a novel gene EOLA1 stimulating ECV304 cell proliferation.
To study the changes in gene expression in endothelial cells stimulated by lipopolysaccharide (LPS) we performed subtraction hybridization on control human umbilical vein endothelial cells (HUVEC) versus HUVEC stimulated by LPS. A novel cDNA, named endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1), was cloned from our differentially expressed EST database of HUVEC cDNA library (GenBank Accession No. ). Computational analysis showed that EOLA1 is 1404bp long, encoding a 158aa, 17.8kDa protein, mapped to chromosome Xq27.4 with 5 exons, 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. Stable transfection of EOLA1 stimulates ECV304 cell proliferation. Our data suggest that the physical interaction of EOLA1 and MT2A may have an important role of cell protection in inflammation reaction.
Liang
Ziwen
Z
Institute of Burn Research, Southwest Hospital, Third Military Medical University, State Key Laboratory of Trauma, Burn and Combined Injury, Chongqing 400038, China. liangzw99@yahoo.com.cn
Yang
Zongcheng
Z
D000595:Amino Acid Sequence; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D042783:Endothelial Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008668:Metallothionein / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D020798:Two-Hybrid System Techniques
C495142:EOLA1 protein, human; D008070:Lipopolysaccharides; C491053:MT2A protein, human; D008565:Membrane Proteins; D011994:Recombinant Proteins; D008668:Metallothionein