SELECTIVE TRANSPORT OF PROTEINS AND FROM THE NUCLEUS

IMPORT:
Some proteins are imported from the cytoplasm to nucleus

• Such proteins are responsible for; genome struture and function.

These proteins include:

 DNA POLYMERASE

 RNA POLYMERASE

 Transcription factors

 Splicing factors

TARGET OF PROTEINS:
Proteins are targeted by specific amino acids sequences called “Nuclear Localization
Signals”

• DISCOVERY OF NLS:

 First nuclear localization signal was characterized by Alan smith and colleagues in
1984.

 These investigators studied Simian virus (SV40) T antigen ( a virus encoded protein
that initiates viral DNA replication).

 Signal responsible for its nuclear localization was identified by:

Finding mutation of single lysine residue, which prevents nuclear import.

T ANTIGEN:
• T antigen nuclear localization sequence consist of seven amino acid sequence.

• ( pro-lys-lys-lys- arg -lys-val)

T anigen NLS diagram

NLS OF NUCLEOPLASMIN:
It has following characteristics:

 Bipartite ( have two parts)

 Having lys-arg sequence followed by a lys-lys-lys sequence located 10 amino acid

further.

 Central part (10 amino acids) mutated ( because no one is as important)

NLS OF RIBOSOMAL PROTEIN:

• Unrelated to basic amino acid rich nls of either T –antigen or nucleoplasmin signals.

TRANSPORTATION:

IMPORT:
has two steps

1st step: Recognition

• Requires no energy

• Proteins having NLS binds to and recognized by receptors.

 • Receptors are called IMPORTIN.

• IMPORTIN SUBUNITS.

o It has two sub units:

o Importin alpha ( binds to basic amino acid rich NLS)

o Importin beta ( binds to cytoplasmic filaments of nuclear pore complex)

o The NLS of ribosomal proteins recognized by distinct receptors.

• These are related to importin beta.

2nd step: Binding and transport:

• Energy dependent process.

• Requires GTP hydrolysis.

• Key player is GTP binding protein called Ran.

Concentration of Ran GTP and Ran GDP

• Enzymes that stimulates GTP binding to Ran are present to nuclear side.

• High concentration of Ran GTP at nuclear side.

• Enzymes that stimulates hydrolysis of GTP are present at cytoplasmic side.

• GTP converts into GDP

• High concentration of Ran GDP at cytosolic side.

BINDING:
• Importin beta forms a complex with alpha and cargo protein.

• Binding on cytoplasmic side . In the presence of high concentration of Ran GDP.

• PROTEIN + importin alpha + importin beta + Ran/GDP = Complex.

TRANSPORTATION:
• Complex transport to nucleus through nuclear pore complex.

• Inside Ran GTP concentration is high.

 • GDP is converted to GTP.

• Ran GTP Binds to importin beta.

• Displace importin alpha and protein.

• Importin alpha and target proteins release in nucleus.

• Importin beta + Ran GTP moves outside the nucleus.

• Because outside Ran GDP is high so, Ran GAP stimulates hydrolysis of bound GTP
into GDP.

• Importin beta releases and participate in another cycle.

IMPORT :

• FIGURE:

EXPORT:
RECOGNITION:

 Proteins that have to be export from nucleus to cytoplasm contains signals.

 These signals are called “ Nuclear Export Signal”

 These signals are recognized by receptors exportin.

 Exportins are related to importin beta.

 Ran GTP promotes the formation of complex

(exportin+proteins+Ran GTP)

EXPORT:
 • This complex is transported to cytoplasm.

• Outside Ran GAP stimulates the hydrolysis of GTP.

• GTP converts into GDP.

• Release exportin and target protein into cytoplasm.

• Exportin is transported again to nucleus.

FIGURE:

DIFFERENCE BETWEEN IMPORT AND EXPORT:

IMPORT EXPORT
• NLS is importin. • NES is exportin.

• Ran GTP dissociates • Ran GTP forms the

the complex complex (exportin +
importin alpha + protein + RanGTP)
importin beta +
protein.