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The plasma membrane (PM) comprises distinct subcellular domains with diverse functions that need to be dynamically coordinated with intracellular events, one of the most impactful being mitosis. The Kv2.1 voltage-gated potassium channel is conditionally localized to large PM clusters that represent specialized PM:endoplasmic reticulum membrane contact sites (PM:ER MCS), and overexpression of Kv2.1 induces more exuberant PM:ER MCS in neurons and in certain heterologous cell types. Localization of Kv2.1 at these contact sites is dynamically regulated by changes in phosphorylation at one or more sites located on its large cytoplasmic C terminus. Here, we show that Kv2.1 expressed in COS-1 cells undergoes dramatic cell cycle-dependent changes in its PM localization, having diffuse localization in interphase cells, and robust clustering during M phase. The mitosis-specific clusters of Kv2.1 are localized to PM:ER MCS, and M phase clustering of Kv2.1 induces more extensive PM:ER MCS. These cell cycle-dependent changes in Kv2.1 localization and the induction of PM:ER MCS are accompanied by increased mitotic Kv2.1 phosphorylation at several C-terminal phosphorylation sites. Phosphorylation of exogenously expressed Kv2.1 is significantly increased upon metaphase arrest in COS-1 and CHO cells, and in a pancreatic β cell line that express endogenous Kv2.1. The M phase clustering of Kv2.1 at PM:ER MCS in COS-1 cells requires the same C-terminal targeting motif needed for conditional Kv2.1 clustering in neurons. The cell cycle-dependent changes in localization and phosphorylation of Kv2.1 were not accompanied by changes in the electrophysiological properties of Kv2.1 expressed in CHO cells. Together, these results provide novel insights into the cell cycle-dependent changes in PM protein localization and phosphorylation.
Pubmed ID: 26442584 RIS Download
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This unknown targets IgG1
View all literature mentionsThis polyclonal secondary targets IgG1
View all literature mentionsThis polyclonal secondary targets IgG2a
View all literature mentionsThis unknown targets Mouse IgG1 (??1)
View all literature mentionsThis monoclonal targets Rat Kv2.1 synthetic phosphopeptide amino acids 595–616, CPEATRFSH[pS]PLASLSSKAGSS, accession NP_037318
View all literature mentionsThis polyclonal secondary targets Rabbit IgG (H+L)
View all literature mentionsThis polyclonal secondary targets Rabbit IgG (H+L)
View all literature mentionsThis unknown targets Mouse IgG2a
View all literature mentionsThis monoclonal targets Rat Kv2.1 synthetic peptide amino acids 581-590 CMSSIDpSFISSA, accession NP_037318
View all literature mentionsThis monoclonal targets Kv2.1
View all literature mentionsThis monoclonal targets Kv2.1 potassium channel
View all literature mentionsThis monoclonal targets Potassium voltage-gated channel subfamily B member 1 (Delayed rectifier potassium channel 1) (DRK1) (Voltage-gated potassium channel subunit Kv2.1)
View all literature mentionsThis monoclonal targets Kv1.4 potassium channel
View all literature mentionsThis monoclonal targets Mortalin/GRP75
View all literature mentionsThis polyclonal targets Rat Kv2.1 synthetic peptide amino acids 710-728, CDKPVLpSPESSIYTTASART, accession NP_037318
View all literature mentionsThis polyclonal targets Rat Kv2.1 synthetic phosphopeptide amino acids 447-467, KDAFAR[pS]IEMMDIVVEKNGES, accession NP_037318, kdafarsiemmdivveknges
View all literature mentionsThis unknown targets phospho-Histone H3 (Ser10)
View all literature mentionsThis polyclonal targets Rat Kv2.1 amino acids 207-225, CLPELQSLDEFGQSTDNPQL , accession NP_037318
View all literature mentionsThis polyclonal targets Kv2.1-pS603
View all literature mentionsThis polyclonal targets Rat Kv2.1 synthetic peptide amino acids 554-574, accession NP_037318
View all literature mentionsThis polyclonal secondary targets IgG1
View all literature mentionsThis polyclonal secondary targets Rabbit IgG (H+L)
View all literature mentionsThis unknown targets Mouse IgG2a
View all literature mentionsThis monoclonal targets Kv2.1
View all literature mentionsThis monoclonal targets Rat Kv2.1 synthetic peptide amino acids 581-590 CMSSIDpSFISSA, accession NP_037318
View all literature mentionsThis monoclonal targets Potassium voltage-gated channel subfamily B member 1 (Delayed rectifier potassium channel 1) (DRK1) (Voltage-gated potassium channel subunit Kv2.1)
View all literature mentionsThis monoclonal targets Kv1.4 potassium channel
View all literature mentionsThis monoclonal targets Mortalin/GRP75
View all literature mentionsThis polyclonal targets Rat Kv2.1 synthetic peptide amino acids 710-728, CDKPVLpSPESSIYTTASART, accession NP_037318
View all literature mentionsThis polyclonal targets Rat Kv2.1 synthetic phosphopeptide amino acids 447-467, KDAFAR[pS]IEMMDIVVEKNGES, accession NP_037318, kdafarsiemmdivveknges
View all literature mentionsThis unknown targets phospho-Histone H3 (Ser10)
View all literature mentionsThis polyclonal targets Rat Kv2.1 amino acids 207-225, CLPELQSLDEFGQSTDNPQL , accession NP_037318
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View all literature mentionsSoftware used for visualizing and graphing data, image processing, and programming. It is designed for use by scientists and engineers and supports large data sets, evenly spaced data, and various data import formats. The software includes a suite of image processing operations for image filtering, manipulation, and quantification and is completely programmable.
View all literature mentionsMulti-channel data acquisition software. Multi-channel stimulation/acquisition software with programmable experiment control and automation, software Lock-in amplifier and Photometry/Imaging extension. PATCHMASTER supports all HEKA amplifiers and data acquisition hardware.
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View all literature mentionsMicroscope imaging software suite used with Nikon products. NIS-Elements includes software applications for advanced and standard research, documentation, confocal microscopy, and high-content analysis.
View all literature mentionsDigital image processing system where microscope settings and processing steps may be adjusted in single user interface. Can acquire images from variety of cameras. Includes software package for capturing, archiving and preparing images for publication. Allows users to visualize and present images in several dimensions. Functionality of imaging toolbox expands constantly with wide range of different modules that are tailored to specific applications or microscope accessories. This resource is duplicated by SCR_018376
View all literature mentionsThis polyclonal secondary targets Rabbit IgG (H+L)
View all literature mentionsThis polyclonal secondary targets Rabbit IgG (H+L)
View all literature mentionsThis unknown targets Mouse IgG2a
View all literature mentionsThis monoclonal targets Rat Kv2.1 synthetic peptide amino acids 581-590 CMSSIDpSFISSA, accession NP_037318
View all literature mentionsThis monoclonal targets Kv2.1
View all literature mentionsThis monoclonal targets Kv2.1 potassium channel
View all literature mentionsThis monoclonal targets Potassium voltage-gated channel subfamily B member 1 (Delayed rectifier potassium channel 1) (DRK1) (Voltage-gated potassium channel subunit Kv2.1)
View all literature mentionsThis monoclonal targets Kv1.4 potassium channel
View all literature mentionsThis monoclonal targets Mortalin/GRP75
View all literature mentionsThis polyclonal targets Rat Kv2.1 synthetic peptide amino acids 710-728, CDKPVLpSPESSIYTTASART, accession NP_037318
View all literature mentionsThis polyclonal targets Kv2.1-pS603
View all literature mentionsThis polyclonal targets Rat Kv2.1 synthetic phosphopeptide amino acids 447-467, KDAFAR[pS]IEMMDIVVEKNGES, accession NP_037318, kdafarsiemmdivveknges
View all literature mentionsThis unknown targets phospho-Histone H3 (Ser10)
View all literature mentionsThis polyclonal targets Rat Kv2.1 amino acids 207-225, CLPELQSLDEFGQSTDNPQL , accession NP_037318
View all literature mentionsThis monoclonal targets Kv2.1
View all literature mentionsThis polyclonal secondary targets IgG1
View all literature mentionsThis polyclonal secondary targets IgG2a
View all literature mentionsThis unknown targets IgG1
View all literature mentionsThis monoclonal targets Kv2.1 potassium channel
View all literature mentionsThis monoclonal targets Kv2.1 potassium channel
View all literature mentionsThis unknown targets Mouse IgG1 (??1)
View all literature mentionsThis polyclonal secondary targets IgG1
View all literature mentionsThis polyclonal secondary targets IgG2a
View all literature mentionsThis unknown targets IgG1
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