Cancer
I. Chromosome and Chromatin
Gene
number 6000 19,000 13,500 30,000 30,000
Simple Calculation
Human beings have roughly 3 billion base pairs
of DNA in 23 pairs of chromosomes
Distance between bases is 3.4 Angstrom
For human this would be 3.4x3x109
Angstroms. or 1.02 meters per haploid
genome,
2.04size
The meters pernucleus
of the cell is roughly 10 micrometers
in diameter
The average size of condensed mitotic human
chromosomes is ~ 1 micrometer (>10,000 fold)
If there is no compaction, nucleus would be too small
To hold all DNA!!!
Function
• Storage of genetic information
• Precise segregation of replicated DNA into two
daughter cells
• Platform for transcription, replication, recombination
and DNA repair
Problem
How to retrieve genetic information from DNA
packaged into chromosomes?
How the long linear DNA molecules are
packaged into compact chromosomes?
Historic View: Chromosomes and
Chromatin
1879 : Walter Flemming discovered chromosomes,
observing threadlike structures in the nuclei
of salamander cells during cell division
Early 1900s, cytologists Walter Sutton and
Theodor Boveri, independently published
papers linking chromosomes to the Mendelian
principles of segregation and independent
assortment.
Their work inspired the chromosomal theory
of inheritance
large rRNA
Compaction of chromatin is cell-stage dependent
A. Interphase chromatin B. a mitotic chromosome,
which is duplicated already
A) 30 nm fibers
B) beads on a stringnucleosome
From interphase nucleus
Composition of Chromatin
DNA
Stable association
Histones (H2A, H2B, H3, H4 and linker histones)
Nonhistone chromosomal proteins
In general not as stable as DNA-histone interactions
Nomenclature
Nucleosome = a nucleosome core particle + linker
DNA+ a linker histone
DNA length: 180-200 bp
Nucleosome core particle = histone octamer + 146 bp DNA
Nucleosomes can be
isolated by digesting
with nucleases that cut
between the nucleosomes
in a region called the linker
Nucleosomeoctamer
2 each H2A, H2B, H3, H4
142 hydrogen bonds between DNA and nucleosome,
mostly between phosphodiester bonds and amino acid
backbone of histones
Histones- highly basic (+) proteins
Core particle
Determining
180-200bp 146 bp sequence
How to determine rotational positioning experimentally?
M phase
Chromosome
scaffold
topoisomerase
Histone depleted
metaphase chromosomes
Scaffold Attachment
Regions (SARs)
• Regions of the chromosomes with
sequences specific for topoisomerase,
HMG protein, and histone H1 binding
• Found only in untranscribed regions of the
eukaryotic chromosomes
• Spaced along the chromosomes, with the
intervening regions containing one or
more genes?
• Highly AT rich (65%) and may be several
hundred bp long
The state of condensation of chromosomes
varies according to the cell growth cycle. The
mitotic chromosomes are highly condensed, in
contrast to that of the interphase chromosome.
What structures are necessary to a
functional eukaryotic chromosome?
• Centromeres
• Telomeres
• Origins of replication
Three types of specialized sequences found in all eucaryotic
chromosomes ensure that chromosomes replicate efficiently.
many, to
ensure speed
kinetochore =
protein
complex that
binds the
spindle and the
centromere
•Chromosome movement
66
– there are two types of heterochromatin:
1. Constitutive heterochromatin, condensed at all times
– includes AT-rich satellite DNA at the centromere
– the telomeres, or ends of the chromosomes (especially in
plants)
2. facultative heterochromatin transient condensation,
contains potentially active genes
– e.g. less active chromosome: chromosome 18
– e.g. one X chromosome (from mother or father) is
“turned off” early in development
» this becomes a “Barr body”
» in subsequent daughter cells, the same chromosome
stays condensed
» eventually the Barr body does get reactivated when
new germ cells are made
– facultative heterochromatin becomes more abundant in
cells as the organism matures from embryo to adult, as
cells specialize fewer genes are active
22.228 lecutre 7 67
Lampbrush and polytene chromosomes
Lampbrush chromosomes: found in the oocytes of many animals.
They contain very transcriptionally active DNA, where loops of DNA
emerging from an apparently continuous chromosomal axis are coated
with RNA polymerase. Each RNA polymerase is attached to nascent
RNA and associated proeins generating a visible ‘brush-like’
appearance.
The transcription of the RNA precursor of the 28S, 18S, and 5.8S ribosomal RNAs. These
units are tandemly linked together, some 450 per haploid genome.
Polytene Chromosomes of Drosophila