NUCLEUS Eukaryotic Nucleus -

Structure

• Largest organelle of the • Major components

cell – An apparent amorphous mass
enclosed by a nuclear envelope
• Contains nearly all DNA – Within:
• RNA synthesis • Chromosomes
– Chromatin
• Varies in shape • Nuclear matrix
• Varies in number within a – Fibrillar network
• Nucleoli
cell – r-RNA and ribosomes
• Nucleoplasm
– Fluid of the nucleus

Topic 4-1 1
 Eukaryotic Nucleus - Structure
• The nuclear envelope
– Components:
• Two parallel cellular membranes
– Perinuclear cisterna
– Barrier to ions, solutes, macromolecules
• Outer membrane
– Ribosomes
– Continuous with RER
• Inner membrane
– Meshwork of fibrous proteins
– With lamins
• Membranes fused to form pores
– Complex assemblies of proteins
 Eukaryotic Nucleus - Structure
• The nuclear envelope
– Components:
 Eukaryotic Nucleus - Structure
• The nuclear pore complex (NPC)
– Huge macromolecular complex
– Nuclear pore + glycoproteins
– 80-100 nm in diameter
– 3 ring-like arrays of protein
– Octagonal symmetry
• 8-fold repetition of subunits
• 30-50 proteins – nucleoporins
– Nucleoporins
• Symmetrical – on both cytoplasmic and nuclear sides
 Eukaryotic Nucleus - Structure
• The nuclear pore complex (NPC)
Eukaryotic Nucleus – Structure/Function
• The nuclear pore complex (NPC)

– Low molecular wt solutes

• Diffuse freely
– Macromolecules
• Regulated
• Mediated by receptors
• Energy-requiring
– “bidirectional” transport mechanism
Eukaryotic Nucleus – Structure/Function
• The nuclear pore complex (NPC)

– Transport receptors
• Soluble
• Importins
– Cytoplasm to nucleus
– Proteins from ribosomes
• Exportins
– Nucleus to cytoplasm
– Transports RNA
Eukaryotic Nucleus – Structure/Function
• Importin
Eukaryotic Nucleus – Structure/Function
• Exportin
CHROMATIN

-interphase

- “uncoiled” chromosome

- guide to the cell’s activity

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 Euchromatin
– The regions of DNA containing active
genes.
• Dispersed, active DNA
• Lighter appearance, located in interior of
nucleus
– L.M. : lightly stained basophilic
– E.M. : beads on string

11
Beads on string

-nucleosomes : beads
-octomeric
-duplicates of 4 types of Histones
 H2A H2B H3 H4

+++++

naked DNA
nucleosome core particle

nucleosomes

Add H1
H1
- 168 bp in loop
H1
- 6 to 7 fold shortening
H1 H1 H1
- 10 nm fiber 13
 Heterochromatin
• Inactive, condensed chromatin
• Located at the edges of the nucleus
– visible by E.M. as a darker colour
– Basophilic clumps under L.M.
• Stains deeply with Feulgen’s stain
 Heterochromatin
• Two types:
– Constitutive heterochromatin is non-coding and
so can never become active
– Facultative heterochromatin contains genes that
are shut off and temporarily inactive. -depends
• In any cell only a portion of all the genes are
active
• Example: the Barr Body. Females have two X
chromosomes, one of which is inactive
 Facultative vs. Constitutive Heterochromatin

Facultative
telomere
Heterochromatin
Euchromatin in
Facultative interior of
Heterochromatin Interphase nucleus
Barr Body inactive X.
nucleus

Euchromatin
Active X-chromosome
Constitutive
Heterochromatin
Telomere
around centromere and
telomere
Cytoplasm

16
 NUCLEOPLASM
• Consists of IGs, PCGs, RNPs + nuclear matrix
• Nuclear matrix: 10% CHONs
30% RNA
1-3% DNA
2-5% PO4-3
 NUCLEOLUS

• Spherical

• Highly basophilic

• Active in protein
synthesis

• Non-membrane bound
structure

• Assembles 40s and 60s

ribosomal subunits
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