Steps at which eukaryotic gene expression can be...
Transcript of Steps at which eukaryotic gene expression can be...
![Page 1: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/1.jpg)
Steps at which eukaryotic gene expression can be controlled
Cell 7.5
![Page 2: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/2.jpg)
Protein Variabilityand
Protein ActivityControl
Aminoacid sequence
Three-dimensional shape
(conformation)
Function
Protein processing
Degradation
![Page 3: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/3.jpg)
Four types of post-translational processingevents
![Page 4: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/4.jpg)
Protein Folding
The aminoacid sequence contains all the
information needed to fold the polypeptide into its
correct tertiary structure
![Page 5: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/5.jpg)
The cellular mechanisms that monitor protein quality after protein synthesis
Cell 6.85
![Page 6: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/6.jpg)
The co-translational folding of a protein
Cell 6.81
The cellular chaperone machinery is specifically recruited to bind to ribosomes and protects nascent chains and folding intermediatesfrom nonproductive interactions
![Page 7: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/7.jpg)
Two main ATP-dependent classes of chaperons, Hsp70 chaperon and cylindrical chaperonin complexes, mediate protein folding
Structure of GroEL/GroES chaperonin(Hsp60)
Hsp70 chaperones bind tohydrophobic regions in unfolded polypeptides, including those thatare still being translated, and holdprotein in an open conformationuntil it is ready to be folded
![Page 8: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/8.jpg)
Protein processing
Proteolytic Cleavage
Chemical Modification
(Intein splicing)
![Page 9: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/9.jpg)
Protein processing by proteolyticcleavage
Genomes 11.27
![Page 10: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/10.jpg)
Ex. proteolytic cleavage: melitin and insulin
Genomes 11.28
Promelittin Melitin
24 aa
22 aa
Extracellular protease
Signal peptide- hydrophobic aa sequence thatattachs preproinsulin to the membrane, beforeexporting the protein through the membrane tothe extracellular environment
A chain C chain B chain110 aa
30 aa21 aa
![Page 11: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/11.jpg)
(51 peptide)
Converting preproinsulin to insulin
![Page 12: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/12.jpg)
Ex. proteolytic cleavage : the pro-opiomelanocortinpolyprotein
POMC is glycosylated and then cleaved to give a numberof neurohormones that can be cleaved enzymaticallyinto the following peptides (neurohormones)
In anterior pituitary gland
In the intermediate lobeof pituitary gland
In the intermediate lobeof pituitary gland- melanin productionandneurons in the arcuatenucleus the hyphotalamus- apetite
Each of these peptides is packaged in large dense-corevesicles that are released from the cells by exocytosisin response to appropriate stimulation
260 amino acid protein
![Page 13: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/13.jpg)
Chemical Modification
![Page 14: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/14.jpg)
Post-translational chemical modificationof calf histone H3
Genomes 11.30
Lysine acetylation and methylation of histone H3
![Page 15: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/15.jpg)
Various carboxylase enzymesLysineBiotinylation
Addition of biotin
Some protein kinases involved in signal transduction
N-terminal glycineN-myristoylation (miristic acid)
Many membrane proteinsSerine, threonine, cysteineAcylation
Addition of lipid side chains
Many membrane proteins and secreted proteins
AsparagineN-linked glycosylation (amino group)
Many membrane proteins and secreted proteins
Serine, threonineO-linked glycosylation (hydroxil groups)
Addition of sugar side chains
MelittinN-terminal glycineN-formylation (COH)
CollagenProline, lysineHydroxylation (OH)
Some proteins involved in signal transduction
Serine, threonine, tyrosinePhosphorylation (P)
HistonesLysineMethylation (CH3)
HistonesLysineAcetylation (CH3CooH)
Addition of small chemical groups
Examples of proteinsAmino acids that are modified
Modification
150 different aa already described
![Page 16: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/16.jpg)
Some ways in which the activity of gene regulatory proteins is regulated in eucaryotic cells
Each of these mechanisms is typically controlled by extracellular signals which are communicatedacross the plasma membrane to the gene regulatory proteins in the cell- SIGNAL TRANSDUCTION
![Page 17: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/17.jpg)
Protein Degradation
![Page 18: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/18.jpg)
Cell 6.82
Protein degradation... when?
![Page 19: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/19.jpg)
Relation between N-terminal amino acid and half-life of E. coli ββββ-galactosidase proteins with modified N-terminal amino acids
N-terminal Amino Acid Half-life
Met, Ser, Ala, Thr, Val, Gly
Ile, Glu
Tyr, Gln
Pro
Phe, Leu, Asp, Lys
Arg
PESTsequence
ProGlnSerThr
Half-life less then 2 hours
more than 20 h
30 min
10 min
7 min
3 min
2 min
![Page 20: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/20.jpg)
Acerca da degradação de proteínas
• Várias vias de degradação – Lisossomas: contêm uma série de hidrolases e enzimas
proteolíticos, degradando essencialmente proteínas transmembranares e do lúmen dos organitos
– Proteossoma: complexo multiproteico que degrada proteínas ubiquitinadas, localizadas sobretudo no núcleo e no citosol.
Ex. Factores de transcrição, proteínas da regulação do ciclo celular como as cinases e fosfatases etc.
- …
• Processos altamente selectivos e rápidos
• Eucariotas- descrito o Proteossoma
![Page 21: Steps at which eukaryotic gene expression can be controlledbmg.fc.ul.pt/Disciplinas/GBM/aulas/15-ProcDegrProt.pdf · Biotinylation Lysine Various carboxylase enzymes Addition of biotin](https://reader034.fdocuments.us/reader034/viewer/2022042312/5edb60a3ad6a402d6665921a/html5/thumbnails/21.jpg)
Lodish 3.13
Ubiquitin and the marking of proteinwith multiubiquitin chains
Ubiquitin-mediatedproteolytic pathway
(7-8 residues)