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JCS ePress online publication date 4 Dec 2002
doi: 10.1242/jcs.00242

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Research Article

CaM kinase II{alpha} mediates norepinephrine-induced translocation of cytosolic phospholipase A2to the nuclear envelope


Soghra Fatima, Fariborz A. Yaghini, Aftab Ahmed, Zinat Khandekar, and Kafait U. Malik*
* Author for correspondence (e-mail: kmalik{at}utmem.edu)

Several growth factors, hormones and neurotransmitters, including norepinephrine, increase cellular calcium levels, promoting the translocation of cytosolic phospholipase A2to the nuclear envelope. This study was conducted to investigate the contributions of the calcium-binding protein calmodulin and of calcium -calmodulin-dependent protein kinase II to cytosolic phospholipase A2translocation to the nuclear envelope elicited by norepinephrine in rabbit aortic smooth-muscle cells. Norepinephrine caused cytosolic phospholipase A2accumulation around the nuclear envelope as determined from its immunofluorescence; cytosolic phospholipase A2translocation was blocked by inhibitors of calmodulin and calcium -calmodulin-dependent protein kinase II or calcium -calmodulin-dependent protein kinase II{alpha} antisense oligonucleotide. Calmodulin and calcium -calmodulin-dependent protein kinase II inhibitors did not prevent cytosolic calcium increase but attenuated cytosolic phospholipase A2phosphorylation caused by norepinephrine or ionomycin. In vascular smooth-muscle cells reversibly permeabilized with {beta}-escin and treated with alkaline phosphatase, norepinephrine failed to cause cytosolic phospholipase A2phosphorylation and translocation to the nuclear envelope; these effects of norepinephrine were minimized by the phosphatase inhibitor okadaic acid. Recombinant cytosolic phospholipase A2phosphorylated by purified calcium -calmodulin-dependent protein kinase II, but not unphosphorylated or dephosphorylated cytosolic phospholipase A2, introduced into permeabilized vascular smooth-muscle cells in the absence of calcium accumulated around the nuclear envelope. These data suggest that norepinephrine-induced translocation of cytosolic phospholipase A2to the nuclear envelope is mediated by its phosphorylation by calcium -calmodulin-dependent protein kinase II and that calcium alone is insufficient for cytosolic phospholipase A2translocation to the nuclear envelope in rabbit vascular smooth-muscle cells.


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