Eukaryotic Genomes
Organization & Regulation
http://www.ncbi.nlm.nih.gov/books/NBK21128/
What is a gene ?
Definitions
1. Classical definition: Portion of a DNA that determines a
single character (phenotype)
2. One gene one enzyme (Beadle & Tatum 1940): Every
gene encodes the information for one enzyme
3. One gene one protein: One gene contains information
for one protein (structural proteins included) one gene
one polypeptide
4. Current definition: A piece of DNA (or in some cases RNA)
that contains the primary sequence to produce a
functional biological gene product (RNA, protein).
Gene classification
intergenic
region
coding genes
non-coding
genes
Chromosome
(simplified)
Messenger RNA
Structural RNA
Proteins
transfer
RNA
Structural proteins
Enzymes
ribosomal
RNA
other
RNA
Cellular Genomes
Viruses Procaryotes
Eucaryotes
Nucleus
Capsid
Plasmids
Viral genome
Bacterial
chromosome
Chromosomes
(Nuclear genome)
Mitochondrial
genome
Chloroplast
genome
Genome: all of an organisms genes plus intergenic DNA
Intergenic DNA = DNA between genes
Prokaryote genomes
Example: E. coli
89% coding
4,285 genes
122 structural RNA genes
Haploid circular genomes
Polycistronic transcription units
Usually asexual reproduction, great variety of
recombination mechanisms
Transcription and translation take place in the same
compartment
Eukaryotic genome
Chromosomes
genes
Coding region
en.wikipedia.org
The Structure of Chromatin
Prokaryotic and eukaryotic cells
Gene expression in eukaryotes has two main differences from
the same process in prokaryotes.
First, the typical multicellular eukaryotic genome is much
larger than that of a bacterium.
Second, cell specialization limits the expression of many
genes to specific cells.
The estimated 35,000 genes in the human genome includes
an enormous amount of DNA that does not program the
synthesis of RNA or protein.
The Structure of Chromatin
Eukaryotic chromosomes contain an enormous amount of DNA
relative to their condensed length.
Each human chromosome averages about 2 x 108
nucleotide pairs.
If extended, each DNA molecule would be about 6 cm
long, thousands of times longer than the cell diameter.
This chromosome and 45 other human chromosomes fit
into the nucleus.
This occurs through an elaborate, multilevel system of
DNA packing.
The Structure of Chromatin
Histone proteins are responsible for the first level of DNA
packaging.
Their positively charged amino acids bind tightly to
negatively charged DNA.
The five types of histones are very similar from one
eukaryote to another and are even present in bacteria.
Unfolded chromatin has the appearance of beads on a
string, a nucleosome, in which DNA winds around a core
of histone proteins.
The Structure of Chromatin
Histones leave the DNA only transiently during
DNA replication.
They stay with the DNA during transcription.
By changing shape and position, nucleosomes
allow RNA-synthesizing polymerases to move
along the DNA.
The beaded string coils to form the 30-nm
chromatin fiber.
This fiber forms looped domains attached to a
scaffold of nonhistone proteins.
Levels of chromatin
packing.
Diagram and transmission
electron micrographs
In a mitotic
chromosome,
the looped domains
coil and fold to
produce the
characteristic
metaphase
chromosome.
These packing steps
are highly specific and
precise with particular
genes
located in the same
places.
Interphase chromatin is generally much less
condensed than the chromatin of mitosis.
The chromatin of each chromosome occupies a
restricted area within the interphase nucleus.
Interphase chromosomes have areas that
remain highly condensed, heterochromatin,
and less compacted areas, euchromatin.
Eukaryotic genes
Most have introns
Produce monocistronic mRNA: only one encoded protein
Introns: intervening sequences within a gene that are not translated
into a protein sequence. Collagen has 50 introns.
Exons: sequences within a gene that encode protein sequences
Splicing: Removal of introns from the mRNA molecule.
Eukaryotic genome
Example: C. Elegans
Transparent nematode (roundworm), about
1 mm in length, which lives in temperate soil
environments. Research into
the molecular and developmental
biology of C. elegans was begun in 1974
by Sydney Brenner and it has since been
used extensively as a model organism
10 chromosomes
19,099 genes
Coding region 27%
Average of 5 introns/gene
Both long and short duplications
en.wikipedia.org
Eukaryotic genome organization
Multiple genomes: nuclear, plastid genomes: mitochondria,
chloroplasts
More about the nuclear genome:
Multiple linear chromosomes, total size 5-10000 MB, 5000 to 50000
genes
Monocistronic transcription units
Discontinuous coding regions (introns and exons)
Large amounts of non-coding DNA
Transcription and translation take place in different compartments
Variety of RNA genes: rRNA, tRNA, snRNA (small nuclear), sno (small
nucleolar), microRNAs, etc.
Often diploid genomes and obligatory sexual reproduction
Standard mechanism of recombination: meiosis
Human Genome Organization
by Dr Finbarr Hayes
HUMAN GENOME
Nuclear genome
3200 Mbp
65-80000 genes
30%
Genes and generelated sequences
Mitochondrial genome
16.6 kb
37 genes
70%
Extragenic
DNA
Two rRNA
genes
Unique or moderately repetitive
10%
90%
Coding
DNA
Pseudogenes
Noncoding
DNA
Gene
fragments
Introns,
untranslated
sequences, etc.
22 tRNA
genes
13 polypeptideencoding genes
80%
20%
Unique or
low copy
number
Moderate to
highly
repetitive
Tandemly
repeated
or clustered
repeats
Interspersed
repeats
Eukaryotic genome
Moderately repetitive
Functional (protein coding, tRNA coding)
Unknown function
SINEs (short interspersed elements)
200-300 bp
100,000 copies
LINEs (long interspersed elements)
1-5 kb
10-10,000 copies
Eukaryotic genome
Highly repetitive
Minisatellites
Repeats of 14-500 bp
1-5 kb long
Scattered throughout genome
Microsatellites
Repeats up to 13 bp
100s of kb long, 106 copies
Around centromere
Telomeres
Short repeats (6 bp)
250-1,000 at ends of chromosomes
Genome Organization at the DNA level
Chromosome organization
Eucaryotic chromosome
Centromere
Telomere
p-arm
Telomere
q-arm
Centromere:
DNA sequence that serve as an attachment for protein during mitosis.
In yeast these sequences (~ 130 nts) are very A+T rich.
In higher eucaryotes centromers are much longer and contain
satellite DNA
Satellite DNA consists of very large arrays of tandemly repeating, non-coding DNA.
Satellite DNA is the main component of functional centromeres, and form the main
structural constituent of heterochromatin
Telomeres:
At the end of chromosomes; help stabilize the chromosome
In yeast telomeres are ~ 100 bp long (imperfect repeats)
Repeats are added by a specific telomerase
Types of DNA repeats
Perfect repeats vs degenerate repeats
Tandem repeats (e.g. satellite DNA)
5-CATGTGCTGAAGGCTATGTGCTGCGACG- 3
3-GTACACGACTTCCGATACACGACGCTGC- 5
Inverted repeats (e.g. in transposons)
5-CATGTGCTGAAGGCTCAGCACATCGACG- 3
3-GTACACGACTTCCGAGTCGTGTAGCTGC- 5
Form stem-loop structures
Palindroms = adjacent inverted repeats
(e.g. restriction sites)
Form hairpin structures
Loop
Stem
Hairpin
Types of DNA sequences in the human genome .
The vast majority of the human genome does not code for human
proteins or RNAs and much of it is repetitive DNA.
Stages in gene expression that can be
regulated in eukaryotic cells.
Colored boxes indicate the processes
most often regulated
A simple model of histone tails and
the effect of histone acetylation .
Histones can also undergo other types of
modifications that help determine
the chromatin configuration.
A eukaryotic gene and its transcript.
Each gene has a promoter, a DNA sequence where RNA polymerase binds ,
a number of control elements, a ploy-A tail and the 5 cap.
A model for the action of enhancers and transcription activators.
Bending of the DNA by a protein enables enhancers to influence
a promoter hundreds or even thousands of nucleotides away.
Cell type-specific transcription.
Both liver cells and lens cells have the genes for
making the proteins albumin and crystallin, but
only liver cells make albumin (a blood protein),
and only lens cells make crystallin (the main
component of the lens of the eye).
Alternative RNA splicing.
The primary transcripts of some genes can be
spliced in more than one way, generating
differente mRNA molecules.
Regulation of gene expression by microRNAs.
Movement of eukaryotic transposable elements.
Transposons are segments of
DNA that can move around to
different positions in the
genome of a single cell.
In the process, they may
cause mutations increase (or
decrease) the amount of DNA
in the genome of the cell, and if
the cell is the precursor of a
gamete, in the genomes of any
descendants.
These mobile segments of
DNA are sometimes
called "jumping genes".
There are two distinct types:
Class II transposons. These
consist of DNA that moves
directly from place to place.
Class I transposons. These
are retrotransposons that
first transcribe the DNA
into RNA and then
use reverse
transcriptase to make a
DNA copy of the RNA to
insert in a new location.
Degradation of a protein by a proteosome.
A proteosome, an enormous protein complex chops up unneeded proteins in the cell.
