Curs 3

Structura Nucleului

Structura nucleului la Eucariote

Componente
Invelisul nuclear Carioplasma (nucleoplasma) - sistem coloidal care include:
Cromozomii Cromatina cu structura fibrilara formata din ADN, proteine nucleare numite histone, ARN, fosfolipide Unul sau mai multi Nucleoli cu rol in sinteza ARN-r si a unor proteine ribozomale
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 Invelisul nuclear
Componente:
doua membrane celulare (intre care se afla spatiul perinuclear) - Membrana interna, sustinuta de lamina nucleara (retea interna de filamente atasate de cromatina) - Membrana externa, in legatura cu RER
Bariera pt. ioni, macromolecule

Membranele formeaza pori

Ansamblu complex de proteine

Eukaryotic Nucleus - Structure

The nuclear envelope
Components:

 Porul nuclear
- Permite transferul unor proteine sintetizate in citoplasma in interiorul nucleului - Transportul in citoplasma a ARN sintetizat in nucleu

 The nuclear pore complex (NPC)

Huge macromolecular complex 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)

Figure 8-72. Freeze-fracture electron micrograph of the elongated nuclear envelope of a fern spore. Note the ordered arrangement of the nuclear pore complexes in parallel rows. In other cells either concentrated clusters of nuclear pores or unusual areas free of nuclear pores have been detected in the nuclear envelope, and these are specifically oriented with respect to othe structures in the cell. (Courtesy of Don H. Northcote; from K. Roberts and D.H. Northcote, 9 Microsc.Acta 71:102-120, 1971.)

Nuclear transport

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Eukaryotic Nucleus Structure/Function

The nuclear pore complex (NPC)
Low molecular wt solutes
Diffuse freely

Macromolecules
Regulated

Protein import to nucleus

Nuclear localization signal (NLS)

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Eukaryotic Nucleus Structure/Function

The nuclear pore complex (NPC)
Transport receptors karyopherins
Soluble Importins
Cytoplasm to nucleus

Exportins
Nucleus to cytoplasm

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Eukaryotic Nucleus Structure/Function

Import example - nucleoplasmin

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Eukaryotic Nucleus Structure/Function

Import example nucleoplasmin
Role of the GTP-binding protein Ran Active form Ran-GTP Inactive form Ran-GDP High concentration of GTP-Ran in nucleus Low concentration in cytoplasm

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Eukaryotic Nucleus Structure/Function

Export RNAs
Move as ribonucleoproteins (RNPs)
Except t-RNA direct transport by exportin-t

Protein component contains nuclear export signal (NES) Exportins recognize NES Binds Ran-GTP stabilizes complex Carried to cytoplasm Ran-GAP1 converts Ran-GTP to Ran-GDP
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Eukaryotic Nucleus Structure/Function

Export RNAs

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DNA second structure

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Cromatina
In Cromatina ADN este legat de proteine numite histone, alcatuite din aminoacizi bazici (Arg, Lys) Histonele formeaza structuri discoidale in jurul carora se infasoara ADN Histonele impreuna cu ADN formeaza unitati repetitive numite NUCLEOZOMI

Nucleosomii se gasesc la intervale de aprox. 200 perechi de perechi de baze de pe moleculele de ADN
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Multiple coiling model

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Basic unit of chromatin-----nucleosomes

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Chromatin type
Euchromatin: open structure active genes Heterochromatin: highly packaged structure inactive genes

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Cromozomii

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Chromosome structure

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Cariotip

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Figure 8-71. Electron micrograph of a mammalian cell nucleus. Note that the condensed chromatin underlying the nuclear envelope is excluded from regions around the nuclear pores. (Courtesy of Larry Gerace.)
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- is

not bounded by a membrane

- Fibrillar center FC which contains DNA that is not being actively transcribed

Nucleolus

- site of rRNA synthesis and which contains RNA molecules ribosome assembly of being synthesized - dynamic stucture, disappear and rebuild in the cell cycle

Dense fibrillar component DFC

in the process

- Granular component which contains maturing ribosomal precursor particles

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Nucleolul

Figure 8-65. Electron micrograph of a thin section of a nucleolus in a human fibroblast, showing its 29 three distinct zones. (A) View of entire nucleus. (B) High-power view of the nucleolus. (Courtesy of E.G. Jordan and J. McGovern.)

 Telomeric DNA
The terminal portion of the telomeres of the chromosomes contains 10-15kb of tandem repeats of a 6bp DNA sequence known as telomeric DNA. The telomeric repeat sequences are necessary for chromosomal integrity in replication and are added to the chromosome by a specialized enzyme known as telomerase. 30

TTGGGG

Replication

Replication

Replication

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Telomerase allows telomere length equilibrium maintenance

Replication

Telomerase

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Telomere shortening leads to cell death

Replication

Replication

Replication

Cell Death

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Many diseases seen in families with insufficient telomerase share features of age-related disease

Bone marrow failure

Immune senescence Chemotherapy intolerance Pulmonary fibrosis

Liver disease
Increased cancer incidence

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