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Identification of a novel human kinesin-related gene (HK2) by the cDNA differential display technique
S Debernardi1, E Fontanella, L De Gregorio
1Division of Experimental Oncology A, National Cancer Institute, Milan, Italy.
Abstract:
We have used the cDNA differential display technique to isolate genes regulated by the synthetic retinoid N-(4-hydroxyphenyl)-all-trans-retinamide (HPR), a cancer chemopreventive agent in vivo and a powerful inducer of apoptotic cell death in vitro. Here we report the identification of a novel gene, the expression of which is markedly up-regulated in tumor cells after treatment for 30-60 min with HPR. The full-length cDNA of this gene, determined by screening of a human placenta cDNA, is 3.5 kb long and contains an open reading frame of 2037 nt. The gene is > 90% homologous to the mouse KIF2, a gene belonging to the family of kinesin-related motor proteins, and we therefore named it HK2 (human kinesin 2). A shorter form of the HK2 mRNA (HK2s), containing a 57-nt deletion in the open reading frame, has also been detected. Northern analysis revealed that HK2 is widely expressed among hemopoietic and nonhemopoietic cell lines and tissues. By the use of radiation hybrids, HK2 has been localized to chromosome 5q12-q13. Kinesins constitute a superfamily of motor proteins that use energy liberated from ATP hydrolysis to move cargo along microtubules and are implicated in mechanisms of mitosis or meiosis. The role of HK2 in the growth-inhibitory and apoptotic responses elicited by HPR remains to be established.
Insights
Researchers identified a novel gene, human kinesin 2 (HK2), upregulated by the cancer chemopreventive agent HPR. This kinesin motor protein
Area of Science:
- Molecular Biology
- Genetics
- Cancer Research
Background:
- Synthetic retinoids like N-(4-hydroxyphenyl)-all-trans-retinamide (HPR) show promise in cancer chemoprevention and induce apoptosis.
- Understanding gene regulation by such agents is crucial for developing targeted therapies.
Purpose of the Study:
- To identify novel genes regulated by HPR.
- To characterize a newly discovered gene involved in HPR-mediated cellular responses.
Main Methods:
- cDNA differential display technique to identify differentially expressed genes.
- Full-length cDNA screening and sequencing.
- Northern blot analysis for gene expression profiling.
- Radiation hybrid mapping for gene localization.
Main Results:
- Identified and characterized a novel gene, named human kinesin 2 (HK2), upregulated by HPR in tumor cells within 30-60 minutes.
- HK2 shares >90% homology with mouse KIF2, a kinesin-related motor protein.
- Detected a shorter HK2 mRNA variant (HK2s) and found HK2 widely expressed across various cell lines and tissues.
- Localized HK2 to chromosome 5q12-q13 using radiation hybrids.
Conclusions:
- HK2 is a novel kinesin-related motor protein gene significantly upregulated by the cancer chemopreventive agent HPR.
- The precise role of HK2 in HPR-induced growth inhibition and apoptosis requires further investigation.