Casein kinase II (CK-II)-mediated stimulation of HIV-1 reverse transcriptase activity and characterization of selective inhibitors in vitro.
The physiological significance of the casein kinase II (CK-II)-mediated phosphorylation of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) on its three enzymatic activities [RNA-dependent DNA polymerase (RDDP), DNA-dependent DNA polymerase (DDDP) and ribonuclease H (RNase H)] was investigated in vitro. It was found that (i) the purified recombinant RT (rRT) functioned as an effective phosphate acceptor for CK-II; (ii) the RDDP, DDDP and RNase H activity of rRT was stimulated about 2.8-, 4.1- and 3.9-fold, respectively, after full phosphorylation by CK-II; and (iii) this stimulation was selectively inhibited by potent CK-II inhibitors, such as neocarzinostatin-chromophore (NCS-chrom) and three polyphenol-containing anti-oxidant compounds [quercetin, epigallocatechin gallate (EGCG) and 8-chloro-3',4',5,7-tetrahydroxyisoflavone (8C-3',4',5,7-THI)]. These results suggest that (i) CK-II may be responsible for activation of RT in HIV-1-infected cells; and (ii) the selective inhibition of CK-II-mediated activation of HIV-1 RT by potent CK-II inhibitors may be involved in the mechanism of their anti-HIV-1 effects at the cellular level.
Harada
S
S
Laboratory of Genetical Biochemistry, School of Allied Health Sciences, Kitasato University, Sagamihara, Japan.
Haneda
E
E
Maekawa
T
T
Morikawa
Y
Y
Funayama
S
S
Nagata
N
N
Ohtsuki
K
K
Nagata
N
N
Ohtsuki
K
K
D019380:Anti-HIV Agents / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D047390:Casein Kinase II; D053281:Enediynes; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004926:Escherichia coli; D006034:Glycyrrhetinic Acid / Q000494:pharmacology; D054303:HIV Reverse Transcriptase / Q000378:metabolism*; D015497:HIV-1 / Q000187:drug effects / Q000201:enzymology*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014162:Transfection; D009353:Zinostatin / Q000031:analogs & derivatives / Q000494:pharmacology
D019380:Anti-HIV Agents; D000975:Antioxidants; D053281:Enediynes; D004791:Enzyme Inhibitors; C032372:neocarzinostatin chromophore; D009353:Zinostatin; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; D054303:HIV Reverse Transcriptase; D006034:Glycyrrhetinic Acid