dna rna protein synthesis amino acids protein protein ... · protein synthesis amino acids. some...
TRANSCRIPT
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DNA
RNA
PROTEIN
FUNCTION
Protein degradation
Protein synthesisAMINO ACIDS
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Some properties of intracellular protein degradation (1970).
• Abnormal proteins are rapidly eliminated.
• Normal proteins are selectively degraded at widely different rates.
• Levels of specific proteins in animal cells can be regulated by
changes in rates of synthesis or rates of degradation (Schimke,
1970).
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Hershko & Tomkins JBC 1971
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Ciechanover, Hod & Hershko, BBRC 1978
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APF-1 (ubiquitin)
Protein (µg) 1.5 3 7 PNAS 1980
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ATPlysozyme (µg)
- -+ + + + +0 0 5 10 25 5 5
125I-ubiquitin
125I-lysozyme
lysozyme- (Ub)n
Hershko et al., PNAS 1980
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PNAS 1980
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DUBDUBDUB
E2E2E2 ProteinProtein
E3E3
UbUb
UbUb
UbUb
UbUb UbUb
UbUb
UbUb
UbUb
UbUb
E2E2E2
E1E1E1
UbUb
E1E1E1
+ATP+ATP
UbUb
AMP+PPAMP+PPiiE1E1E1
E2E2E2
UbUb
++ ATPATP
ADP+PADP+Pii
ProteinProteindegradeddegraded
26S proteasome26S proteasome
UbUb
ProteinProtein
UCHUCHUCH
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Hershko et al., JBC 1983
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Substrate Yes No Yes No Yes Yes
E3α-125I-proteincross-link
125I-protein
Cytochrome c (yeast)
Cytochrome c (mammalian)
Lysozyme
Ribonuclease A (native)
Ribonuclease A (oxidized 1)
Ribonuclease A (oxidized 2)
DETECTION OF E3-SUBSTRATE BINDING BY CHEMICAL CROSS-LINKING
Free 125I-proteinsJBC 1986
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MBC 1995
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Cyclosome/APC
MBC 1995
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UU
UU
UU
UU
Cdk1
Cyclin B
Cdk1
Cyclin B
Cdk1
~ ~~ ~ Cdc20
PP
InactiveCdk1
InactiveAPC/cyclosome
ActiveAPC/cyclosome
E1, E2-CATP, Ub(b)
ATP
(a)
Cyclinsynthesis
26SProteasome
ActiveCdk1
Phosphatase
(d)
Cyclindegraded
Pi
PP
Cdc20
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APC/Cinactive
APC/Cactive
APC/Cinactive
APC/C = Anaphase-PromotingComplex/Cyclosome
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G0/G1 TimeTime
Lev
elL
evel
S
p27Cyclin E Cyclin A
Skp2
p27Cyclin E
Cdk2
Cyclin E
Cdk2
Cyclin A
Cdk2
Cdk2 inactive Cdk2 activep27 degraded
Cks1
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NCB 1999
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NCB 1999
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The three binding sites of Cks1
Cdk binding(green)Anion binding
(blue)
Skp2 binding(red)
JBC 2002
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p27Cyclin E
Cdk2p27
Cyclin E
Cdk2
Cyclin E
Cdk2
Cdk2 inactive
p27-(Ub)n
P
p27Cyclin E
Cdk2
Skp1
SCF complex
P
Skp2 &Cks1
synthesis
Skp2Cks1
Cyclin Esynthesis
Cdk2 active
p27 degraded
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Some roles of ubiquitin-mediated protein degradation.
• Control of cell division• Signal transduction• Regulation of gene expression• Responses to inflammation• Immune response• Embryonic development• Apoptosis• Circadian clocks
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Some regulatory proteins degraded by the ubiquitin system.
Regulator Role ofdegradation
Examples
I Positive Limitation of duration
Cyclins (G1, S, M-phase); transcription factors (myc, fos…)
II Negative Initiation of process
Cdk inhibitors Anaphase inhibitorIκB transcriptional regulator
III Positive or negative
Activation by stabilization
p53 tumor suppressor; β-catenin
Type
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2000
Microbe hunters
Vitaminhunters
Enzyme hunters
Gene hunters
1900
Arthur Kornberg : For the Love of Enzymes (1989)
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Technion lab (1971-present)
Dvora GanothHanna HellerEsther EytanSarah EliasClara SegalJudith Hershko
Former graduate studentsAaron CiechanoverYuval ReissValery SudakinShirly Lahavand many others…
Collaboration and helpIrwin A. RoseLeonard CohenJoan RudermanMichele Pagano
Present graduate studentsGil BornsteinDanielle Sitry-ShevahYakir Moshe
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