Transport in and out of the nucleus

LODISH 6E
• The nuclear envelope consists of two phospholipid membranes
that are continuous with the ER membrane.

• Nuclear pores span the inner and outer nuclear membranes.

• Nuclear pores are made up of a structure called the nuclear

pore complex (NPC).

• The NPC is 60-80 MD and is 16X larger than a ribosome.

• NPC consists of multiple copies of ~30 different proteins called

nucleoporins.
Octagonal shape of NPC as seen by SEM
Model of NPC

FG nucleoporins
 NPC
• Nuclear basket made up of 8 nuclear filaments with a
terminal ring in nucleoplasm.
• Membrane portion of the basket is attached to nuclear
lamina (lamins).
• Cytoplasmic filaments extend from the cytoplasmic face of
the NPC into the cytoplasm.
• Proteins that function in the nucleus (histones; DNA and RNA polymerases;
TFs) contain NLS.
• Most NLS are similar consisting of 7 basic AA sequence located near the C-
terminal end of the protein.
• Por-Lys-Lys-Lys-Arg-Lys-Val (prototype NLS of large T-antigen of SV40).
• Some NLS are chemically different consisting mainly of hydrophobic AAs.
NLS of SV40 large T-antigen
Cytoplasmic proteins leak out into the
medium (lysate)
Digitonin-treated HeLa cells incubated with a fluorescent labelled protein
chemically coupled to a synthetic SV40 NLS. Such experiments lead to the
purification of three proteins (Ran + importin-alpha + importin-beta).
• Ran:
– A small GTPase protein.

• Importin-alpha and –beta: can form a hetrodimeric nuclear import

receptor .
– Alpha; binds to NLS on cargo proteins.
– Beta; binds to proteins found in the NPC; beta may also bind to certain
NLS and therefore function independently as a nuclear import
receptor.
Nuclear import by importins

B-nucleoporin specifically interacts with FG-nucleoporins which

line the channel of the NPC and are also found in the nuclear
basket and cytoplasmic filaments; FG proteins are rich in
phenylalanine and glycine.
In the nucleoplasm, the importin interacts with Ran-GTP, this
interaction leads to release of cargo protein from importin.
Exportin interacts with FG-nucleoporins.
 Karyopherins family

• Importins and exportins; are very similar in structure, they belong the
to same family.
• ~ 14 karyopherins in yeast and ~20 in mammals.
• The NLS and NES to which they bind have not all been
charactrrized.
• Some karyopherins can function as importins or exportins.

Export and import occur by simple diffusion down concentration

gradients and interactions with FG-nucleoporins.