Lecture 3 - Regulation Of Chromatin Structure Flashcards
What are centromeres characterised by?
Specific histone H3 variant (CenpA or CenH3)?
What do centromeres in higher eukaryotic chromosomes contain in large amounts?
Repetitive DNA and unique histone variants
What type of structure does a centromere have?
Heterochromatin
What is the structure of heterochromatin like?
The structure is inaccessible and repressive No Replication No Transcription No DNA repair
What is the structure of Euchromatin like?
The structure is accessible and extended Replication Transcription DNA repair
What is chromatin remodelling?
The energy-dependent displacement of reorganisation of nucleosomes that occurs in conjugation with activation of genes for transcription There are numerous ATP-dependent chromatin remodelling dimpled that use energy provided by hydrolysis of ATP
What can remodelling complexes do?
Alter, slide or displace nucleosomes
What are histone post translational modification?
Covalent in nature Lysine methylation Lysine acetylation Lysine ubiquitilation Lysine sumolyation
Arginine methylation Glutamate ADP ribosylation Threonine phosphorylation.
Serine phosphorylation Tyrosine phosphorylation Arginine deimination (citrullination)
What is side chain modification?
Histone tails are highly conserved and are modified on specific amino acids Modification of amino acid side chains can alter structure/function
What is direct effect of side chain modification?
Change in charge
What is the indirect effect of side chain modification?
Change in charge
Histone modification
Histone modifications are mutually exclusive e.g. lysine cannot be methylated if already acetylated and vice versa Marks are reversible
What does chromatin modification function depends on?
Specific modification ‘reader’
Methylation
KMTs
Demethylation
KDMs
Acetylation
HDACs
De acetylation
HATs
Phosphorylation
Kinase
Dephosphorylation
Phosphatases
What is HAT?
Histone acetyltransferase
What is Lysine acetylation/deacetylation of histone tails?
Acetylation of Lysine residues in histone tails Loss of positive charge Reduction in binding to DNA Opening up of chromatin Access to DNA
HDAC
Histone deacetylase
What is TAFII250 bromodomain?
Acetylates Transcriptional activation
Histone modifying complexes have several components
Histone modification enzymes have little, if any, specificity other than for Lysine, they have the ability to acetylation/deaceytlate ANY Lysine on ANY histone or nucleosome
What is histone acetylation associated with?
Transcription activation
What is Lysine (K) acetyltransferase (KAT) ?
An enzyme present in large complexes that acetylates Lysine residues in histones AKA histone acetyltransferase
What is transcription activators associated with?
Histone acetylase activities in large complexes
What is histone deacetylase (HDAC)?
Enzyme that removes acetyl groups from histones May be associated with repressors of transcription
Where are deacetylase present ?
Complexes with repressor activity
Where do distinct enzyme activities exist ?
Both methylation for and demethylating Lysines (mono, di, tri)
What is context dependent?
Different methylated Lysine’s can recruit activators or repressors
What is methylation catalysed by?
Histone methyltransferase (KMTs)
What is demethylation catalysed by?
Histone demethyltransferase (KDMs)
What is HP1 chromodomain?
Methylates Heterochromatin assembly Transcriptional silencing
How can processes such as transcription be regulated?
By switching modifications
What is DNA methylation?
Covalent addition of a methyl group onto 5th position of cytosine within self complementary CpG dinucleotides Catalysed by DNA methyltransferase
What does DNA methyltransferase do ?
Flip bases out of the double helix
What are examples of highly methylated sequences?
Satellite DNA Repetitive element (transposons and relic) Non-repetitive intergenic DNA and exons of genes
What is the percentage of CpG in human somatic cells that are methylated (global methylation)?
70-80%
Approximately how many CpG islands in human genome?
45,000
Where are CpG islands present?
Nearly all constitutively active house keeping genes
What are CpG islands in normal tissue?
Unmethylated
What does DNA methylation do?
Compact chromatin and is associated with Transcriptional repression
What are the 3 types of modification that affect chromatin?
- Acetyltransferase/deacetylase 2. Histone methyltransferase/demethylase 3. DNA demethylase/ DNA methyltransferase
Acetyltransferase/deacetylase
Remodelling complexes can facilitate binding of acetyltransferase complexes
Histone methyltransferase/demethylase
Histone methylation can also recruit chromatin-modifying complexes
What do different modifications and complexes facilitate?
Transcription, initiation and elongation
DNA demethylase/ DNA methyltransferase
DNA methylation and histone modification act in concert to regulate transcription in the chromatin environment
What are modification?
Reversible and therefore dynamic
Inactive to active state
Histone acetyltransferase Histone demethylase DNA demethylase
Active to inactive state
Histone deacetylase Histone methyltransferase DNA methyltransferase