Effect of HIV integrase inhibitors on the RAG1/2 recombinase.
Assembly of functional Ig and T cell receptor genes by V(D)J recombination depends on site-specific cleavage of chromosomal DNA by the RAG1/2 recombinase. As RAG1/2 action has mechanistic similarities to DNA transposases and integrases such as HIV-1 integrase, we sought to determine how integrase inhibitors of the diketo acid type would affect the various activities of RAG1/2. Both of the inhibitors we tested interfered with DNA cleavage and disintegration activities of RAG1/2, apparently by disrupting interaction with the DNA motifs bound specifically by the recombinase. The inhibitors did not ablate RAG1/2's transposition activity or capture of nonspecific transpositional target DNA, suggesting this DNA occupies a site on the recombinase different from that used for specific binding. These results further underscore the similarities between RAG1/2 and integrase and suggest that certain integrase inhibitors may have the potential to interfere with aspects of B and T cell development.
Melek
Meni
M
Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Building 5, Room 241, Bethesda, MD 20892, USA.
Jones
Jessica M
JM
O'Dea
Mary H
MH
Pais
Godwin
G
Burke
Terrence R
TR
Pommier
Yves
Y
Neamati
Nouri
N
Gellert
Martin
M
D020816:Amino Acid Motifs; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002474:Cell-Free System; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D019428:HIV Integrase Inhibitors / Q000494:pharmacology*; D018398:Homeodomain Proteins / Q000378:metabolism*; D008274:Magnesium / Q000494:pharmacology; D051379:Mice; D008954:Models, Biological; D011485:Protein Binding; D011995:Recombination, Genetic; D013997:Time Factors
D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; D019428:HIV Integrase Inhibitors; D018398:Homeodomain Proteins; C499308:Rag2 protein, mouse; C064655:V(D)J recombination activating protein 2; C064658:RAG-1 protein; D008274:Magnesium; D002118:Calcium