the nucleus text and image sources are included using the notes function of the.ppt file
TRANSCRIPT
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The Nucleus
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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 HistoneCode
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 reconstru
ction
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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NuclearmRNA
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
factorsidentifies
nucleoporins on a
Western-style blot
Nup358Nup214Nup153
Nup 98
Nup 62
Nuclear Transport
Ran*GDP
Ran*GTP
Ran*GDP
Ran*GTP
Cytoplasmic compartment Nuclear compartment
Ran-GEFRan-GAP
Exportin
NESExportin
NES
NLS ImportinNLS Importin
FG affinity transport
Follow the FG-repeatsFollow the FG-repeatsFollow, follow, follow, follow
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Animation
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The nucleus - not just a place to find DNA
anymore