- Authors: Kallio, H., Tolvanen, M., Janis, J., Pan, P. W., Laurila, E., Kallioniemi, A., Kilpinen, S., Tuominen, V. J., Isola, J., Valjakka, J., Pastorekova, S., Pastorek, J., Parkkila, S.
- Year: 2011
- Journal: PLoS ONE 6 e27152
- Applications: in vitro / siRNA / INTERFERin
- Cell types:
- Name: HeLa
Description: Human cervix epitheloid carcinoma cells
- Name: SU.86.86
Description: Human ductal pancreas carcinoma
- Name: U-373 MG
Description: Human glioblastoma-astrocytoma, grade III
Known as: U-373
- Name: HeLa
Our previous microarray study showed that the non-specific cytotoxic cell receptor protein 1 (Nccrp1) transcript is significantly upregulated in the gastric mucosa of carbonic anhydrase IX (CA IX)-deficient (Car9(-/-)) mice. In this paper, we aimed to characterize human NCCRP1 and to elucidate its relationship to CA IX. Recombinant NCCRP1 protein was expressed in Escherichia coli, and a novel polyclonal antiserum was raised against the purified full-length protein. Immunocytochemistry showed that NCCRP1 is expressed intracellularly, even though it has previously been described as a transmembrane protein. Using bioinformatic analyses, we identified orthologs of NCCRP1 in 35 vertebrate genomes, and up to five paralogs per genome. These paralogs are FBXO genes whose protein products are components of the E3 ubiquitin ligase complexes. NCCRP1 proteins have no signal peptides or transmembrane domains. NCCRP1 has mainly been studied in fish and was thought to be responsible for the cytolytic function of nonspecific cytotoxic cells (NCCs). Our analyses showed that in humans, NCCRP1 mRNA is expressed in tissues containing squamous epithelium, whereas it shows a more ubiquitous tissue expression pattern in mice. Neither human nor mouse NCCRP1 expression is specific to immune tissues. Silencing CA9 using siRNAs did not affect NCCRP1 levels, indicating that its expression is not directly regulated by CA9. Interestingly, silencing NCCRP1 caused a statistically significant decrease in the growth of HeLa cells. These studies provide ample evidence that the current name, "non-specific cytotoxic cell receptor protein 1," is not appropriate. We therefore propose that the gene name be changed to FBXO50.