Beruflich Dokumente
Kultur Dokumente
90 nm
FG Repeat/non-
structure
Nucleoporins
FG Repeat
Binding Site
FG Repeat
Binding Site
Nucleoporins: glycosylated &
hydrophobic
• Mammals • Yeasts
– FXFG – GLFG
H3
Phosphorylation Me Ac
H3
Ubiquitin as a sorting Signal
GH receptor Endocytosis
EGF receptor internalization
Glutamate receptor
etc
Proteasome In Eukaryotes
Threonine Proteases
Beta1, beta2, beta5 with catalytic activity.
Beta1: post-glytamyl-peptide-hydrolyzing; beta2:
trypsin like; beta5:chymotrypsin like.
catalytic
Beta 1, 2 & 5
Regulatory Factors for Proteasome
Assembly
• Positive • Negative
– Hsp90 – PAAF-1 (proteasomal
ATPase-associated
factor-1)
Closed Open
Activator & Inhibitor
19S=PA700
PA28
PI31
PI31
Phosphorylation
19S
• Base • Lid: Non-ATPase
– Rpt1-Rpt6: ATPase – 8 subunits
– Non-ATPase – Rpn10: a hinge between
• Rpn1 the base and lid
• Rpn2 – Rpn11: An intrinsic Zn2+-
dependent
metallopeptidase to release
ubiquitin and peptides for
20S proteasome
– Rpn4: Binds and acts as a
transcriptional activator
PA28 (Proteasome Activators)
• Alpha/beta • Gamma
– In immune tissues – For trypsin-like activity
– cytoplasmic • In brain
• In nucleus
Immuno-Proteasomes
• Page 43
Fool the substrates for E3
RING
Virus
HECT
E3
RING
• RING (Really Interesting New Gene)
– Two zinc atoms
– No directly associated with ubiquitin
K
48
K
63
Ubiquitin
E3
• SCF (Skip-Cullin-F- • Anaphase-Promoting
box) Complex (APC;
– Phosphorylation of he cyclosome) contains
target protein often a RING-finger protein
triggers ubiquitylation (Apc11), an E2 and a
cullin (Cdc20/Hct1).
SCF
RING: Roc1
ECS Substrate
binding
HIF: Hypoxia
Inducing Factor
E3: Cell Cycle Regulatory
Anaphase-
promoting
complex
Skp-Cul1-
F-box: with
Phosphorylated
substrates
Phosphorylation
Protein Folding versus Degradation
• CHIP (C-terminal Hsp70-interacting
protein) contains a modified RING-finger
and promotes the ubiquitylation of
misfolded proteins, which are then
degraded by the proteasome.
Ubiquitin like protein
Ubiquitin-Like
Protein
Protein
Protein
Export
Crm1
Aster promoting activity
Leucine-rich Nuclear Export
Signals
• Low affinity for CRM1, stimulated by
RanBP2
• CRM1 –mediated translocation through
the NPC is energy-independent facilitated
diffusion
Sumoylation at Nuclear Pore
Complex
RanBP2(Nup358)
Models
Small Ubiquitin-like Modifier
(SUMO)
SUMO
•97 aa
•SUMO-1, -2, -3, -4
•SUMO modification occurs on the lysine in
ψKXE
Ψ:hydrophobic
Substrates for Sumoylation
• RanGAP ψKXE/D
• PCNA TKET
• Smad4 VKYC
• VKDE
E3 ubiquitin ligase
mRNA export
NXF1-p15 (Mex67p:Mtr2p)
mediate nuclear export of bulk
mRNA
THO: tetrameric
protein complex
TREX: couple
transcription &
export
RNA binding
domain
Cell-Cycle Regulation
M-Phase-Promoting Factor
Mitotic spindle
Activation of M-Phase-Promoting
Factor
References
• Journal of Biological Chemistry
280(38)32565-32568
• Journal of Biological Chemistry
279(52)53899-53902
• Essays in Biochemistry: The Ubiquitin-
Proteasome System (edited by RJ Mayer
&R Layfield)
• zonnc@sinica.edu.tw