Molecular cloning and characterization of the human ASB-8 gene encoding a novel member of ankyrin repeat and SOCS box containing protein family.
We have cloned a new member of human ankyrin repeat and SOCS box containing protein family (ASB), designed as hASB-8, from a human placental cDNA library and further extended by 5(') and 3(')-RACE. The full-length cDNA was 2545bp in length, with a predicted open reading frame encoding a protein of 288 amino acids, which was 96% identical to mouse ASB-8 protein. Computer analysis revealed that the deduced amino acid sequence of the human ASB-8 contained four Ankyrin repeats and one SOCS box. The gene had four exons separated by three introns and was mapped to human chromosome 12q13. Human ASB-8 mRNA was expressed at the highest level of expression in skeletal muscle and at a varied level of expression in heart, brain, placenta, liver, kidney, and pancreas. The transcript of hASB-8 was not detected in adult normal lung tissue, but found in lung carcinoma cell lines SPC-A1, A549, and NCI-H446. Subcellular localization analysis showed that the EGFP-tagged hASB-8 protein was localized at cytoplasm in human hepatocellular carcinoma cell line BEL-7402. We also provided evidence that hASB-8 could interact with Elongin B-C complex in vitro. Furthermore, transfection with the truncated mutant of hASB-8 cDNA lacking SOCS box could suppress cell growth of lung adenocarcinoma SPC-A1 cells in vitro, which suggests that this gene might be related to the development of lung cancer.
Liu
Yongzhong
Y
State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Shanghai 200032, PR China.
Li
Jinjun
J
Zhang
Fengrui
F
Qin
Wenxin
W
Yao
Genfu
G
He
Xianghuo
X
Xue
Peng
P
Ge
Chao
C
Wan
Dafang
D
Gu
Jianren
J
D000328:Adult; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D017487:Ankyrins / Q000737:chemistry; D001483:Base Sequence; D002881:Chromosomes, Human, Pair 12; D003001:Cloning, Molecular; D000075065:Elongin; D005091:Exons; D006801:Humans; D007438:Introns; D051379:Mice; D008969:Molecular Sequence Data; D010920:Placenta / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020449:Repetitive Sequences, Amino Acid; D016415:Sequence Alignment; D050826:Suppressor of Cytokine Signaling Proteins; D014018:Tissue Distribution; D014157:Transcription Factors / Q000378:metabolism; D014407:Tumor Cells, Cultured
C472010:ASB8 protein, human; D017487:Ankyrins; C000616396:ELOB protein, human; C000616401:Elob protein, mouse; D000075065:Elongin; D011506:Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D050826:Suppressor of Cytokine Signaling Proteins; D014157:Transcription Factors