How is gene expression in eukaryotes accomplished ?

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Presentation transcript:

How is gene expression in eukaryotes accomplished ?

The control of eukaryotic gene expression: an overview In the nucleus: 1) DNA unpacking involving a) DNA demethylation b) histone acetylation 2) Transcription control 3) primary mRNA transcript processing 4) preparing mRNA for transport

The control of eukaryotic gene expression: an overview In the cytoplasm: 1) Translation controls 2) Degradation of mRNA 3) Post-translational Modification a) cleavage of long polypeptides b) tagging and chemical modification c) transport to cellular destinations d) activating proteins 4) Degradation of proteins

Chromatin organization Condensed heterochromatin is not expressed. A gene’s location relative to nucleosomes and nuclear membrane (scaffold) influences its expression

DNA Demethylation DNA methylation = addition of methyl groups (CH 3 ) after DNA synthesis Usually cytosine can become methylated Genes that are not expressed are more heavily methylated Once methylated, genes usually stay that way through later divisions. Demethylating certain inactive genes turns them on.

Histone Acetylation Acetylation enzymes attach acetyl groups (COCH 3 ) to certain amino acids of histone proteins. Acetylated histones bind to DNA less tightly; transcription factors have easier access to genes

Transcription Control Transcription factors are necessary for RNA polymerase to bind to DNA during transcription. Transcription factors must be able to bind to DNA (DNA-binding domain) and to proteins (protein binding domain) Similar transcription factors activate or repress groups of genes in synchrony.

Transcription Control Eukaryotic genes have the following structural organization: Promoter – RNA polymerase binding Proximal control elements – transcription factor binding sites near promoter Enhancers (distal control elements) – DNA that bind proteins called activators at sites very remote from the promoter Silencers (distal control elements) – DNA that binds proteins called repressors at sites not to far from enhancers

Transcription Control (Hormone Signaling) Steroid (fat-soluble) hormone diffuses through the cell membrane and nucleus. Steroid binds to inactive receptor protein and activates it. Active receptor molecule attaches to specific sites within the enhancer. Enhancer, now active, can bind to activator protein.

Transcription Control How eukaryotic genes are transcribed: 1) Activator proteins bind to enhancer sites on DNA (or repressor proteins bind to silencer sites near enhancer sites and inhibit transcription). 2) DNA bending brings bound activators closer to other transcription factors. 3) Protein-binding domains on the activators attach to transcription factors and help form an active transcription complex 4) RNA polymerase is now free to bind and move along the DNA.

Primary mRNA Transcript Processing and Preparation for RNA Transport Introns must be removed and exons must be spliced. Alternative RNA splicing can occur as exons are arranged in various ways. A 5’ cap and a poly- A tail are added.

mRNA Degradation Eukaryotic mRNA can exist for long periods of times (hours to weeks) example – mRNA for hemoglobin mRNA is degraded when: Poly-A tail is hydrolyzed. 5’ cap is removed. (mRNA codes for this) Nucleases hydrolyze the remaining mRNA molecule from 5’ end.

Control of Translation mRNA is stopped from initiating translation by: Binding translation repressor protein to 5’ end of a mRNA to prevent ribosome attachment Inactivating certain initiation factors Occurs in early embryonic development Egg has stored inactive mRNA prior to fertilization New cells respond with a burst of protein synthesis after fertilization

Protein Processing and Degradation o Many eukaryotic polypeptides must be modified or transported before becoming active. o Modifications include adding phosphates, cleaving large polypeptides, tagging with sugar, marking for export

Protein Processing and Degradation Selective degradation occurs when;  Ubiquitin is added to mark for destruction.  Proteosomes, huge protein hydrolyzing complexes, recognize ubiquitin and degrade the tagged protein  Dangerous exception to the above are mutated cell cycle proteins that proteosomes can not recognize. Cancer may result.