Calcium, ATP and nuclear pore channel gating

被引:36
作者
Bustamante, JO
Michelette, ERF
Geibel, JP
Dean, DA
Hanover, JA
McDonnell, TJ
机构
[1] Univ Tiradents, Nucl Physiol Lab, BR-49037610 Aracaju, Sergipe, Brazil
[2] Yale Univ, Sch Med, Dept Med, New Haven, CT 06520 USA
[3] Yale Univ, Sch Med, Dept Surg, New Haven, CT 06520 USA
[4] Yale Univ, Sch Med, Dept Cellular & Mol Physiol, New Haven, CT 06520 USA
[5] Univ S Alabama, Coll Med, Dept Microbiol & Immunol, Mobile, AL 36688 USA
[6] NIDDK, Lab Cell Biochem, NIH, Bethesda, MD 20892 USA
[7] Univ Texas, MD Anderson Canc Ctr, Dept Mol Pathol, Houston, TX 77030 USA
来源
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY | 2000年 / 439卷 / 04期
关键词
ATP; calcium; calcium ion channels; cancer cells; Dunning G prostate; gene activity; gene expression; gene regulation; nuclear; nuclear envelope; nuclear pores;
D O I
10.1007/s004240050960
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
Nuclear envelope (NE) cisternal Ca2+ and cytosolic ATP are required for nuclear-pore-complex (NPC-) mediated transport of DNAs, RNAs, transcription factors and other large molecules. Isolated cardiomyocyte nuclei, capable of macromolecular transport (MMT), have intrinsic NPC ion channel behavior. The large ion conductance (gamma) activity of the NPC channel (NPCC) is blocked by the NPC monoclonal antibody mAb414, known to block MMT, and is also silenced during periods of MMT. In cardiomyocytes, neither cytosolic Ca2+ nor ATP alone directly affects NPCC gating. To test the role of Ca2+ and ATP in NPCC activity, we carried our the present patch-clamp study with the pipette attached to the outer NE membrane of nuclei isolated from cultured Dunning G prostate cancer cells. Our investigations demonstrate that in these isolated nuclei neither cytosolic Ca2+ nor ATP alone directly affects NPCC gating. However, when simultaneously applied to the bath and pipette, they transiently silence NPCC activity through stimulation of MMT by raising the Ca2+ concentration in the NE cisterna ([Ca2+](NE)). Our fluorescence microscopy observations with nuclear-targeted macromolecular fluorochromes (B-phycoerythrin and plasmid for the enhanced green fluorescence protein EGFP, pEGFP-C1) and with FITC-labeled RNA support the view that channel silence accompanies MMT. Repeated Ca2+ loading of the NE with Ca2+ and ATP, after unloading with 1-5 mu M inositol 1,4,5-trisphosphate (IP3), thapsigargin (TSG) or 5 mM BAPTA or EGTA, failed to affect channel gating. This result indicates that other factors are involved in this phenomenon and that they are exhausted during the first cycle of NE Ca2+ loading/unloading - in agreement with current theories of NPC-mediated MMT. The results explain how Ca2+ and IF, waves may convert the NE into an effective Ca2+ barrier and, consequently, affect the regulation of gene activity and expression through their feedback on MMT and NPCC gating. Thus, [Ca2+](NE) regulation by intracellular messengers is an effective mechanism for synchronizing gene activity and expression to the cellular rhythm.
引用
收藏
页码:433 / 444
页数:12
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