J. Ellenberg et J. Lippincott-schwartz, Dynamics and mobility of nuclear envelope proteins in interphase and mitotic cells revealed by green fluorescent protein chimeras, METHODS, 19(3), 1999, pp. 362-372
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 i
n nuclear organization and dynamics. To address these issues we have attach
ed green fluorescent protein (GFP) to a well-characterized protein of the i
nner nuclear membrane, lamin B receptor, believed to be one of the major ch
romatin docking protein in the NE. We have used this construct in a variety
of applications, including dual-color GFP time-lapse imaging, to investiga
te the mechanisms underlying protein targeting to the NE and NE breakdown a
nd reassembly during mitosis. In this review, we present a summary of the r
esults from such studies and discuss the photobleaching and imaging methodo
logy on which they were derived. (C) 1999 Academic Press.