Isolation of a candidate gene, CAB1, for cholesterol transport to mitochondria from the c-ERBB-2 amplicon by a modified cDNA selection method.
An improved cDNA selection method was established to isolate expressed genes efficiently from an amplified chromosome region in human cancer. Biotinylated yeast artificial chromosome DNA containing c-ERBB-2 was hybridized in solution with PCR-amplifiable cDNAs of an esophageal cancer cell line bearing the c-ERBB-2 amplification. After capturing the hybrids on avidin-coated magnetic beads, the cDNAs were amplified by PCR. Four new genes (A39, C51, CAB1, and GRB-7) coamplified with c-ERBB-2 were isolated from the enriched cDNA library. CAB1, GRB-7, and c-ERBB-2 were overexpressed in gastric and esophageal cancer cells in correspondence with the amplification. The deduced amino acid sequence of the CAB1 gene had significant homology to the recently discovered steroidogenic acute regulatory protein, StAR, which plays an essential role in cholesterol transport to mitochondria. It was established that multiple overexpressed genes are frequently present in a single amplicon.
Akiyama
N
N
Genetics Division, National Cancer Center Research Institute, Tokyo, Japan.
Sasaki
H
H
Ishizuka
T
T
Kishi
T
T
Sakamoto
H
H
Onda
M
M
Hirai
H
H
Yazaki
Y
Y
Sugimura
T
T
Terada
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism*; D002874:Chromosome Mapping; D018076:DNA, Complementary / Q000235:genetics; D004938:Esophageal Neoplasms / Q000235:genetics* / Q000378:metabolism; D051382:GRB7 Adaptor Protein; D018734:Genes, erbB-2 / Q000235:genetics*; D006801:Humans; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013274:Stomach Neoplasms / Q000235:genetics* / Q000378:metabolism; D014407:Tumor Cells, Cultured
D002352:Carrier Proteins; D018076:DNA, Complementary; C495955:GRB7 protein, human; D008565:Membrane Proteins; D011506:Proteins; C105684:STARD3 protein, human; D051382:GRB7 Adaptor Protein; D002784:Cholesterol