Involvement of HMGB1 and HMGB2 proteins in exogenous DNA integration reaction into the genome of HeLa S3 cells.
High mobility group 1 and 2 proteins (HMGB1 and HMGB2) are abundant chromosomal proteins in eukaryotic cells. We examined the involvement of HMGB1 and HMGB2 in nonhomologous illegitimate recombination. The HMGB1 or HMGB2 expression plasmid, carrying the neo(r) gene as a selection marker, was introduced into HeLa S3 cells to obtain stably-transfected cells. The number of G418-resistant colonies was about 10 times the number of colonies of control cells transfected with plasmids not carrying the HMGB genes. The copy number of the stably-integrated neo(r) gene was higher in the cells transfected with the HMGB expression plasmids than in control cells. The exogenous DNA integration was suggested to have occurred by nonhomologous illegitimate recombination. On the contrary, the introduction of the HMGB antisense RNA expression plasmid with a reporter plasmid carrying the neo(r) gene into HeLa S3 cells decreased the number of G418-resistant colonies. These results indicate that HMGB1 and HMGB2 each have a novel function as stimulators of stable integration of plasmid DNA into the host genome and that they may be important for the process of spontaneous DNA integration in living cells.
Ueda
Tetsuya
T
Department of Biological Science and Technology, Science University of Tokyo, 2641 Yamazaki, Noda, Chiba 278-8510, Japan.
Shirakawa
Hitoshi
H
Yoshida
Michiteru
M
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D004351:Drug Resistance / Q000235:genetics; D018014:Gene Transfer Techniques; D005839:Gentamicins / Q000494:pharmacology; D024243:HMGB1 Protein / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D024261:HMGB2 Protein / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D019868:Kanamycin Kinase / Q000235:genetics; D010957:Plasmids / Q000235:genetics / Q000378:metabolism*; D011995:Recombination, Genetic / Q000502:physiology*
D000900:Anti-Bacterial Agents; D005839:Gentamicins; D024243:HMGB1 Protein; D024261:HMGB2 Protein; C010680:antibiotic G 418; D019868:Kanamycin Kinase