Zinc stimulates Mg2+-dependent 3'-processing activity of human immunodeficiency virus type 1 integrase in vitro.
Human immunodeficiency virus type 1 integrase (HIV-1 IN) catalyzes both 3'-donor processing and strand transfer reactions. Previous studies have determined that the N-terminal region, a putative zinc finger, is capable of binding Zn2+. The function of zinc coordination to this domain, however, is still unknown. In this report, we present evidence that Mg2+-dependent 3'-donor processing by HIV-1 IN is enhanced by the addition of Zn2+ in vitro. This activity is inhibited in the presence of the chelator 1,10-phenanthroline (OP). In addition, the Mg2+-dependent 3'-donor processing activity is more sensitive to the concentration of IN than is the Mn2+-dependent activity. A combination of dimethyl sulfoxide (DMSO) and poly(ethylene glycol) (PEG) was found to further activate the Mg2+-dependent 3'-donor processing activity while diminishing the Mn2+-dependent activity. These results suggest factors such as substrate-length, concentration of IN, Zn2+ coordination, and protein-protein interactions are important for efficient and specific donor processing activity with Mg2+ in vitro.
Lee
S P
SP
Department of Biochemistry and Molecular Biology, Georgetown University Medical Center, Washington, D.C. 20007, USA.
Han
M K
MK
D001483:Base Sequence; D004254:DNA Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics; D015497:HIV-1 / Q000187:drug effects / Q000201:enzymology*; D019426:Integrases; D008274:Magnesium / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism; D010618:Phenanthrolines / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D015032:Zinc / Q000494:pharmacology*
D004268:DNA-Binding Proteins; D009838:Oligodeoxyribonucleotides; D010618:Phenanthrolines; D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D004254:DNA Nucleotidyltransferases; D019426:Integrases; D008274:Magnesium; D015032:Zinc; C025205:1,10-phenanthroline; D004121:Dimethyl Sulfoxide