Dynamics and mobility of nuclear envelope proteins in interphase and mitotic cells revealed by green fluorescent protein chimeras.
Understanding how membrane proteins are targeted to and retained within the nuclear envelope (NE) and the fate of these proteins during NE disassembly/reassembly in mitosis is central for insight into the function of the NE in nuclear organization and dynamics. To address these issues we have attached green fluorescent protein (GFP) to a well-characterized protein of the inner nuclear membrane, lamin B receptor, believed to be one of the major chromatin docking protein in the NE. We have used this construct in a variety of applications, including dual-color GFP time-lapse imaging, to investigate the mechanisms underlying protein targeting to the NE and NE breakdown and reassembly during mitosis. In this review, we present a summary of the results from such studies and discuss the photobleaching and imaging methodology on which they were derived.
Ellenberg
J
J
Gene Expression and Cell Biology/Biophysics Programmes, EMBL, Menehofstrasse 1, Heidelberg, D-69117, Germany. Jan.Ellenberg@EMBL-Heidelberg.de
Lippincott-Schwartz
J
J
D000818:Animals; D019556:COS Cells; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents / Q000493:pharmacokinetics*; D007399:Interphase / Q000502:physiology*; D008164:Luminescent Proteins / Q000235:genetics / Q000493:pharmacokinetics*; D008938:Mitosis / Q000502:physiology; D008967:Molecular Biology / Q000379:methods*; D009685:Nuclear Envelope / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000493:pharmacokinetics
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins