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Cloning, chromosomal localization, and tissue expression of autotaxin from human teratocarcinoma cells
1Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Abstract:
Autotaxin, a potent human tumor cell motility-stimulating exophosphodiesterase, was isolated and cloned from the human teratocarcinoma cell line NTera2D1. The deduced amino acid sequence for the teratocarcinoma autotaxin has 94% identity to the melanoma-derived protein, 90% identity to rat brain phosphodiesterase I/nucleotide pyrophosphatase (PD-I alpha), and 44% identity to the plasma cell membrane marker PC-I. Utilizing polymerase chain reaction screening of the CEPH YAC library, we localized the autotaxin gene to human chromosome 8q23-24. Northern blot analysis of relative mRNA from multiple human tissues revealed that autotaxin mRNA steady state expression is most abundant in brain, placenta, ovary, and small intestine.
Insights
Researchers isolated and cloned autotaxin, a protein that stimulates tumor cell movement, from human teratocarcinoma cells. The autotaxin gene was located on chromosome 8, with high mRNA expression found in the brain and placenta.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Autotaxin (ATX) is a secreted enzyme known to stimulate tumor cell motility.
- Understanding the genetic basis and tissue distribution of ATX is crucial for cancer research.
Purpose of the Study:
- To isolate and clone human autotaxin from a teratocarcinoma cell line.
- To determine the chromosomal localization of the autotaxin gene.
- To analyze the tissue-specific expression of autotaxin mRNA.
Main Methods:
- Autotaxin was isolated and cloned from the human teratocarcinoma cell line NTera2D1.
- Polymerase chain reaction (PCR) screening of a human YAC library was used for gene localization.
- Northern blot analysis was performed to assess mRNA expression in various human tissues.
Main Results:
- The deduced amino acid sequence of teratocarcinoma autotaxin showed high identity to melanoma-derived autotaxin and rat brain phosphodiesterase I/nucleotide pyrophosphatase (PD-I alpha).
- The autotaxin gene was mapped to human chromosome 8q23-24.
- Autotaxin mRNA was most abundant in the brain, placenta, ovary, and small intestine.
Conclusions:
- Human autotaxin has been successfully isolated and cloned, providing a basis for further functional studies.
- The localization of the autotaxin gene to chromosome 8 offers insights into its genetic regulation.
- The widespread expression of autotaxin in various tissues, particularly the brain, suggests diverse physiological roles beyond tumor cell motility.