Cloning and characterization of the human novel gene encoding 6.2 kDa protein highly expressed upon ultraviolet irradiation in DNA-PK-deleted human glioma M059J cells.
The full length of a gene isolated using a methylated DNA binding column as previously reported (Cross et al., 1994) has been cloned. The gene encodes 55 amino acids (6.2 kDa), and has a transcript of approximately 0.45 kb. It has 2 exons in the genome. After searching for sequence homology (BLAST), a homology to hypothetically 6.3 kDa protein located at cosmid ZK652 (protein P34660; EMBL Accession No. L14429) of C. elegans chromosome 3 was noted (51.9% amino acid homology). It has 1 site (-SQ-) for DNA-dependent protein kinase (DNA-PK) phosphorylation and 2 sites of myristilation. This novel gene is very strongly expressed (about 25.6 times) in DNA-PK-deleted human glioma cell line (M059J) 2 hrs after ultraviolet (UV) irradiation compared to the DNA-PK +/+ cell line (M059K) which expresses the gene at a relatively low level (about 7.5 times). Therefore, this result suggests that DNA-PK may negatively regulate the transcriptional level of this gene in M059K cells upon UV damage (50 J/m2), associated with cell toxicity.
Lee
J S
JS
Institute for Molecular Biology and Genetics, Seoul National University, South Korea. jslee2@plazal.snu.ac.kr
D000595:Amino Acid Sequence; D001483:Base Sequence; D002470:Cell Survival / Q000528:radiation effects; D016543:Central Nervous System Neoplasms / Q000235:genetics* / Q000532:radiotherapy*; D003001:Cloning, Molecular; D004249:DNA Damage / Q000528:radiation effects; D019175:DNA Methylation; D051747:DNA-Activated Protein Kinase; D004268:DNA-Binding Proteins; D004307:Dose-Response Relationship, Radiation; D015972:Gene Expression Regulation, Neoplastic / Q000528:radiation effects; D005910:Glioma / Q000201:enzymology / Q000235:genetics* / Q000532:radiotherapy*; D015801:Helminth Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D011506:Proteins / Q000235:genetics* / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured; D014466:Ultraviolet Rays
D004268:DNA-Binding Proteins; D015801:Helminth Proteins; D008565:Membrane Proteins; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D024101:Mitochondrial Proteins; D009687:Nuclear Proteins; D011506:Proteins; C405562:TOMM7 protein, human; D051747:DNA-Activated Protein Kinase; C497263:PRKDC protein, human; D017346:Protein Serine-Threonine Kinases