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I principali compartimenti intracellulari di una cellula animale
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Sequenze e Zone SEGNALE
dirigono le proteine al corretto indirizzo cellulare
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THE NUCLEUS Euchromatin (“normal”)
05_19_Interphase.jpg CHROMATIN Euchromatin (“normal”) Heterochromatin (highly condensed, ~10%)
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Nucleus Structure About 10% of the cell volume
Contains DNA - condensed and organized with proteins as chromatin Nuclear matrix – protein-containing fibrillar network Nucleolus - (pl. nucleoli) –rRNA synthesis, ribosome assembly Surrounded by nuclear envelope
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Active chromatin (euchromatin) appear lighter
Inactive chromatin (heterochromatin) is darker and located near periphery of nucleus Nucleolus a granular region site of ribosome assembly Interphase nucleus
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Nuclear envelope (INVOLUCRO NUCLEARE):
2 lipid bilayer membranes, nm apart the outer membrane is continuous with endoplasmic reticulum a denser layer of intermediate filaments, the nuclear lamina, is within the inner membrane composed of lamin proteins provides structure to nucleus, heterochromatin binding site
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12 year boy with progeria Defect in lamin A gene results in progeria – premature aging
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Il complesso del poro nucleare
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Nuclear Pores: ~3000/nucleus how the cell moves molecules into and out of the nucleus.
At the pore, the inner and outer membranes come together forming an opening. Opening is lined with proteins: nuclear pore complex at least 30 different nucleoporin proteins, in octagonal symmetry ( total proteins) Including cytoplasmic filaments, extending to the cytoplasm
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The role of nuclear pores
Regulate transport of RNA and proteins into and out of nucleus Traffic is FAST! 1 typical human cell has to import 560,000 ribosomal proteins/minute, export 14,000 assembled ribosomes/minute Small molecules and proteins of less than 9 nm can pass freely Large proteins must be assisted Nuclear pores act as turnstiles
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Nuclear Localization Signal (NLS)
-Pro-Pro-Lys-Lys-Lys-Arg-Lys-Val-
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What happens when we alter the NLS?
Kalderson, Roberts, Richardson, Smith (1984) Cell 39:p499. mutation
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Another Experiment Microinjection Pipettes
What happens when we use recombinant DNA techniques to add the NLS to a dummy protein? Normal or modified Bovine Serum Albumin (NLS added) and injected to the cytoplasm Microinjection Pipettes Normal BSA BSA with NLS
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So What? This provided evidence of nuclear transport receptors
family of proteins associated with the nuclear pore complex Importins recognize the NLS and bring proteins in Another set of proteins, the exportins, work in the opposite direction (These recognize other signals)
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15_09_pore_transport.jpg NLS 15_09_pore_transport.jpg
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L’involucro nucleare è disassemblato durante la mitosi
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