5-Lipoxygenase compartmentalization in granulocytic cells is modulated by an internal bipartite nuclear localizing sequence and nuclear factor kappa B complex formation.
A region of basic amino acids spanning residues 639-656 in the human 5-lipoxygenase sequence resembles a consensus bipartite nuclear localizing sequence. A synthetic peptide consisting of the Kaposi fibroblast growth factor signal sequence fused to the 5-lipoxygenase639-656 bipartite nuclear localizing sequence has a prominent inhibitory effect on 5-lipoxygenase catalysis in granulocytic HL-60 cells activated by calcium ionophor A23187. Recombinant 5-lipoxygenase was not affected by the peptide. The peptide also inhibited redistribution of 5-lipoxygenase from the cytosol to the nuclear membrane of HL-60 cells stimulated by A23187. 5-Lipoxygenase protein was detected in nuclear factor kappaB (NF-kappaB) p65 subunit immunoprecipitate fractions prepared from HL-60 cell lysates. The amount of 5-lipoxygenase protein coimmunoprecipitated by NF-kappaB antiserum was increased following A23187 stimulation. In cells treated with agents that block 5-lipoxygenase translocation to the nucleus, 5-lipoxygenase protein appearing in the NF-kappaB immunoprecipitate was diminished. Our results implicate an internal bipartite nuclear localizing sequence as a regulatory domain that modulates 5-lipoxygenase redistribution and catalysis in granulocytic cells. Additionally, our results suggest that molecular determinants which govern 5-lipoxygenase and NF-kappaB redistribution to the nucleus may be coordinately controlled in granulocytic cells.
Lepley
R A
RA
Department of Pharmacology, University of Colorado Health Sciences Center, Denver, Colorado, 80262, USA. Robert.Lepley@UCHSC.edu
Fitzpatrick
F A
FA
D000595:Amino Acid Sequence; D001094:Arachidonate 5-Lipoxygenase / Q000187:drug effects / Q000378:metabolism*; D002451:Cell Compartmentation / Q000187:drug effects; D004789:Enzyme Activation / Q000187:drug effects; D005419:Flavonoids / Q000494:pharmacology; D006098:Granulocytes / Q000187:drug effects / Q000201:enzymology*; D018922:HL-60 Cells; D006801:Humans; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D019913:Nuclear Localization Signals / Q000187:drug effects / Q000502:physiology*; D010446:Peptide Fragments / Q000494:pharmacology; D011233:Precipitin Tests; D013347:Subcellular Fractions / Q000201:enzymology
D005419:Flavonoids; D016328:NF-kappa B; D019913:Nuclear Localization Signals; D010446:Peptide Fragments; D001094:Arachidonate 5-Lipoxygenase; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one