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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 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