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Page 1: The Nucleus Text and image sources are included using the notes function of the.ppt file

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The Nucleus

Text and image sources are included using the notes function of the .ppt file

Page 2: 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

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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

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The HistoneCode

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Anchoring the genome

LAPs (Lamin associated proteins) anchor the lamina to the inner envelope

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Freeze etch view of the lamina

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The nuclear matrix

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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

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Total RNA production (fluorouridine pulse)

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Nucleolar Substructure

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Mammalian cell

Yeast cell

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Polymerase I transcribes

rDNA in nucleolar fibrillar centers

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500 nm

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Gene Expression Factories

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Animated reconstru

ction

Euchromatin (acH3), DNA (DAPI), Spliceosomal protein SC-35

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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)

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Nuclear traffic

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Visualizing

Nuclear Pores

Negative Stain

Thin Section

Freeze Fracture

CryoEM Tomographic Reconstruction

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Annulate lamellae store pores

400 nm

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Nuclear pores

In vitro regulation by Ca++

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Transport of nucleoplasmin-coated gold particles

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It takes two (or three) to tango

Digitonin-permeabilized cells incubated with NLS*BSA*TRITC

-cytosol +cytosol +cytosol, GTP

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NLS*BSA + cytosolic

factorsidentifies

nucleoporins on a

Western-style blot

Nup358Nup214Nup153

Nup 98

Nup 62

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Nuclear Transport

Ran*GDP

Ran*GTP

Ran*GDP

Ran*GTP

Cytoplasmic compartment Nuclear compartment

Ran-GEFRan-GAP

Exportin

NESExportin

NES

NLS ImportinNLS Importin

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FG affinity transport

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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