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The Nucleus
Text and image sources are included using the notes function of the .ppt file
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Interphase Nuclear Dynamics
DIC Brightfield Euchromatin
Time Lapse Video
EM
thin
section
view
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Chromatin organization
early replication -> late replication chromosomal organization one cell cycle later
lamin B
and
late
replicated
DNA
lamin B
and
early
replicated
DNA
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Packing
it all in
146 bp; 6x condensation
40x condensation
The Histone
Code
Anchoring the genome
LAPs
(Lamin
associated
proteins)
anchor the
lamina to
the inner
envelope
Freeze etch view of the lamina
The nuclear matrix
Diversity of nuclear shapes
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Eosinophil nuclei are constricted
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Chromosomal domains
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Nuclear compartments
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Promyelocytic leukemia (PML)
nuclear bodies
Video
Total RNA production
(fluorouridine pulse)
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Nucleolar Substructure
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Mammalian cell
Yeast cell
Polymerase I transcribes
rDNA in nucleolar fibrillar centers
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500 nm
Gene Expression Factories
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Animated
reconstruction
Euchromatin (acH3), DNA (DAPI), Spliceosomal protein SC-35
Spliceosomes
contain
poly-A RNA
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Nuclear speckle
dynamics
Time lapse sequences
show SC-35 protein
over 30 min periods
3 Video Sequences
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Nuclear
mRNA
localizations
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Collagen A1 mRNA
Epstein-Barr Viral mRNAs
mRNA (red)
Gene (green)
Nuclear traffic
Visualizing
Nuclear
Pores
Negative Stain
Thin Section
Freeze
Fracture
CryoEM Tomographic Reconstruction
Annulate lamellae store pores
400 nm
Nuclear
pores
In vitro
regulation
by Ca++
Transport of
nucleoplasmin-
coated gold
particles
It takes two (or three) to tango
Digitonin-permeabilized cells incubated with NLS*BSA*TRITC
-cytosol +cytosol +cytosol, GTP
NLS*BSA
+ cytosolic factors
identifies
nucleoporins on a
Western-style blot
Nup358
Nup214
Nup153
Nup 98
Nup 62
Nuclear Transport
Ran*GDP
Ran*GTP
Ran*GDP
Ran*GTP
Cytoplasmic compartment Nuclear compartment
Ran-GEF Ran-GAP
Exportin NES Exportin NES
NLS Importin NLS Importin
FG affinity transport
Follow the FG-repeats Follow the FG-repeats Follow, follow, follow, follow
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Animation
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The nucleus - not just a place to
find DNA anymore