The document discusses genome organization in eukaryotes. It begins by defining the genome as an organism's entire hereditary information, encoded in DNA or RNA. In eukaryotes, DNA is associated with histone proteins to form chromatin fibers, which condense into chromosomes. The document then discusses various levels of chromatin organization, from DNA wrapping around nucleosomes to form beads on a string, to higher-order folding forming metaphase chromosomes. Chromatin exists in two types - loosely packed euchromatin and tightly packed heterochromatin. Overall, the document provides an overview of eukaryotic genome and chromatin organization from nucleosomes to chromosomes.
Basics of Undergraduate/university fellows
Transcription is more complicated in eukaryotes than in prokaryotes because
eukaryotes possess three different classes of RNA polymerases and because of the
way in which transcripts are processed to their functional forms.
More proteins and transcription factors are involved in eukaryotic transcription.
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
Basics of Undergraduate/university fellows
Transcription is more complicated in eukaryotes than in prokaryotes because
eukaryotes possess three different classes of RNA polymerases and because of the
way in which transcripts are processed to their functional forms.
More proteins and transcription factors are involved in eukaryotic transcription.
BAC & YAC are artificially prepared chromosomes to clone DNA sequences.yeast artificial chromosome is capable of carrying upto 1000 kbp of inserted DNA sequence
Ribonucleic acid (RNA) is a polymeric molecule essential in various biological roles in coding, decoding, regulation, and expression of genes. RNA and DNA are nucleic acids, and, along with proteins and carbohydrates, constitute the four major macromolecules essential for all known forms of life. Like DNA, RNA is assembled as a chain of nucleotides, but unlike DNA it is more often found in nature as a single-strand folded onto itself, rather than a paired double-strand.
What is Genome ?
Types of Genome
Genetic Organization
Genome organization in prokaryotes
BACTERIAL GENOME
Importance of Plasmid
Packaging of DNA
Genome organization in eukaryotes
Chemical composition of chromatin
Nucleosome model
Prokaryotic Genome v/s Eukaryotic Genome
Prokaryotic and eukaryotic gene structurestusharamodugu
Organization of genome in Prokaryotes:
The term prokaryote means “primitive nucleus”. Cell in prokaryotes have no nucleus. The prokaryotic chromosome is dispersed within the cell and is not enclosed by a separate membrane. Much of the information about the structure of DNA comes from studies of prokaryotes, because they are less complex than eukaryotes. Prokaryotes are monoploids they have only one set of genes (one copy of the genome). In most viruses and prokaryotes, the single set of genes is stored in a single chromosome (single molecule either RNA or DNA).
Organization of genome in Prokaryotes:
The term prokaryote means “primitive nucleus”. Cell in prokaryotes have no nucleus. The prokaryotic chromosome is dispersed within the cell and is not enclosed by a separate membrane. Much of the information about the structure of DNA comes from studies of prokaryotes, because they are less complex than eukaryotes. Prokaryotes are monoploids they have only one set of genes (one copy of the genome). In most viruses and prokaryotes, the single set of genes is stored in a single chromosome (single molecule either RNA or DNA). Organization of genome in Prokaryotes:
The term prokaryote means “primitive nucleus”. Cell in prokaryotes have no nucleus. The prokaryotic chromosome is dispersed within the cell and is not enclosed by a separate membrane. Much of the information about the structure of DNA comes from studies of prokaryotes, because they are less complex than eukaryotes. Prokaryotes are monoploids they have only one set of genes (one copy of the genome). In most viruses and prokaryotes, the single set of genes is stored in a single chromosome (single molecule either RNA or DNA).
Differences in DNA occur within genes, the differences have the potential to affect the function of the
gene and hence the phenotype of the individual. Genetic markers which have been used a lot in the past
include blood groups and polymorphic enzymes. We have relatively few such markers, but this has been
overcome with the advent of new types of markers. However, most molecular markers are not associated
with a visible phenotype.
Molecular biology is the study of biology at a molecular level.
In broad sense, the study of gene structure and functions at the molecular level to understand the molecular basis of hereditary, genetic variation, and the expression patterns of genes.The field overlaps with other areas of biology and chemistry, particularly genetics and biochemistry.
Chromatin is the complex combination of DNA and proteins that makes up chromosomes. It can be made visible by staining with specific techniques and stain (thus the name chromatin which literally means colored material). The major proteins involved in chromatin are histone proteins; although many other chromosomal proteins have prominent roles too. The functions of chromatin is to package DNA into smaller volume to fit in the cell, to strengthen the DNA to allow mitosis and meiosis and to serve as a mechanism to control gene expression and DNA replication.
Silkworm has developed an efficient host defense mechanism against invading microorganisms through their immunological and genetic resistance. Immunological responses in silkworm B.mori are provided by circulating haemocytes which play an important role in innate immune mechanism such as phagocytosis, cellular encapsulation, phenoloxidase cascade and synthesis of antimicrobial proteins which are effectively engaged in defense reactions against invading pathogens . Antimicrobial proteins are the armament that insects have developed to fight off the pathogens. Several such antimicrobial proteins have been reported from silkworm B.mori like cecropins, attacins, lebocin, moricin, gloverins, lysozyme, defensins and hemolin. Insects in general are observed to respond differentially to infection by pathogens. Such differences are genetically determined and have been extensively studied in silkworm to develop resistant breeds.
2. What is Genome ?
Genome is the entirety of an organism's hereditary
information.
It is encoded either in DNA or, for many types of virus, in
RNA.
The genome includes both the genes and the non-coding
sequences of the DNA.
3. „‟Genome is thus the entire collection of genes and all
other functional and non functional DNA sequences in
an organism in a haploid set of chromosomes. It
includes structural genes, regulatory genes and non
functional nucleotide sequences ‟‟
Structural genes-
DNA segments that code for some
specific RNAs or proteins. Encode for mRNAs, tRNAs, snRNAs,
scRNAs.
Functional sequences-
Regulatory sequences- occur
as regulatory elements (initiation sites, promoter sites,operator
sites,etc.)
Nonfunctional sequences-
Introns and repititive
sequences. Needed for coding, regulation and replication of
DNA. Much more in no than functional sequences.
4. Genome of all living organisms represents
their hereditary material and is formed of
DNA.
In Prokaryotic cellsGenomic DNA forms a single circular
chromosome, without basic proteins , lies in the cell
cytoplasm in nucleoid region
In Eukaryotic cellsDNA is associated with basic
proteins(histones), form long chromatin fibers.
Chromatin fibers form a network, enclosed in a
double layered nuclear envelop, condenses into
chromosomes during cell division
5. comparative genome sizes of organisms
Size (bp)
gene
number
average gene density
chromosome
number
Homo sapiens
(human)
3.2 billion
~25,000
1 gene /100,000 bases
46
Mus musculus
(mouse)
2.6 billion
~25,000
1 gene /100,000 bases
40
Drosophila
melanogaster
(fruit fly)
137 million
13,000
1 gene / 9,000 bases
8
Arabidopsis thaliana
(plant)
100 million
25,000
1 gene / 4000 bases
10
Caenorhabditis elegans
(roundworm)
97 million
19,000
1 gene / 5000 bases
12
Saccharomyces
cerevisiae
(yeast)
12.1 million
6000
1 gene / 2000 bases
32
Escherichia coli
(bacteria)
4.6 million
3200
1 gene / 1400 bases
1
H. influenzae
(bacteria)
1.8 million
1700
1 gene /1000 bases
1
organism
6. EUKARYOTIC GENOME
‘The nucleus is heart of the cell, which serves as the
main distinguishing feature of the eukaryotic cells. It is an
organelle submerged in its sea of turbulent cytoplasm which
has the genetic information encoding the past history and
future prospects of the cell. Nucleus contains many thread
like coiled structures which remain suspended in the
nucleoplasm which are known as chromatin substance’
Chromatin is the complex combination of DNA and proteins that
makes up chromosomes.
The major proteins involved in chromatin are histone proteins;
although many other chromosomal proteins have prominent roles
too.
.
The functions of chromatin is to package DNA into smaller
volume to fit in the cell, to strengthen the DNA to allow mitosis and
meiosis and to serve as a mechanism to control gene expression and
DNA replication.
7.
8. ORGANIZATION OF CHROMATIN
In resting non-dividing eukaryotic cells, the genome is in the form
of nucleoprotein-complex- the chromatin.
(randomly dispersed in the nuclear matrix as interwoven network of fine chromatin threads)
The information stored in DNA is organized, replicated and
read with the help of a variety of DNA-binding proteins:
Structural Proteins- Histones(Packing proteins):
main structural proteins found in eukaryotic cells
low molecular weight basic proteins with high proportion of
positively charged amino acids,
bound to DNA along most of its length,
the positive charge helps histones to bind to DNA and play a crucial
role in packing of long DNA molecules.
Functional Proteins- Non- Histones:
associated with gene regulation and other functions of chromatin.
10. oChemical composition of chromatin
DNA= 20-40 %- most important chemical constituent of chromatin
RNA=05-10 %-associated with chromatin as;
Ribosomal RNA-( rRNA)
Messenger RNA- (mRNA)
Transfer RNA- (tRNA)
PROTEINS=55-60%-associated with chromatin as;
Histones: very basic proteins, constitute about 60% of total protein,
almost 1:1 ratio with DNA.
FIVE TYPES: H1, H2a, H2b, H3 and H4
Non-Histones: They are 20% of total chromatin protein:
50% structural (actin, L & B tubulin and myosin)-contractile proteins,
function during chromosome condensation and in the movement of
chromosomes during mitosis and meiosis.
50% include all enzymes and co-factors –involved in replication,
transcription and regulation of transcription.
11. Ultrastructure and organization
Multi-strand model
Folded fiber model
Nucleosome model-( R.D.Kornberg & O. Thomas-1974
widely accepted)
NUCLEOSOME MODEL: DNA is tightly bound to histone
proteins which serve to form a repeating array of DNAProtein particles called nucleosomes.
THREE LEVELS OF ORGANIZATION:
DNA wrapping around “Nucleosomes”- The string on beads structure.
A 30 nm condensed chromatin fiber consisting of nucleosome arrays in
their most compact form- The solenoid structure.
Higher levels of packing into metaphase chromosome- The loops, domains
and scaffold structure.
14. Nucleosome Structure
Nucleosomes contain 2 copies
of H2A, H2B, H3 and H4
147 bp of DNA is wrapped
around nucleosome
Histone tails emanate from core
Some nucleosomes
contain histone variants
H1 is a linker histone
from Jiang and Pugh, Nature Rev.Genet. 10, 161 (2009)
15. Hierarchy of Chromatin Organization in the Cell Nucleus:
Nuclear Matrix Associated Chromatin Loops
16.
17. TYPES OF CHROMATIN
Euchromatin
Lightly packed form of chromatin that is rich in gene concentration
takes up light stain and represent most of the chromatin, that disperse after mitosis
has completed.
Consists of structural genes which replicate and transcribe during G1 and S phase
of the interphase.
Considered genetically active chromatin, since it has a role in their phenotypic
expression of the genes.
DNA is found packed in 3-8 mm fibre.
During metaphase it takes up dark stain.
Heterochromatin
Tightly packed form of chromatin that takes up deep stain during interphase and
prophase but metaphase takes up light stain.
Chromomeres, centromeric regions, and knobs also take up dark staining, of which
centromeric regions and knobs are the true Heterochromatic. (chromomeres are
transcribed so not true H.C.).
IN the chromosomes all the centromeres fuse to form a long Heterochromatic mass
called chromocentre.
Heterochromatin consists of highly repetitive DNA sequences. It is late replicating
during the s-phase of the cell and is not transcribed.
19. • Eukaryotic Chromosomes Demonstrate Complex
Organization Characterized by Repetitive DNA
• Repetitive DNA sequences are repeated many
times within eukaryotic chromosomes.
20.
21. • Satellite DNA is highly repetitive and consists
of short repeated sequences.
22. Centromeres
• are the primary constrictions along
eukaryotic chromosomes
• mediate chromosomal migration during
mitosis and meiosis
23. • Telomeric DNA sequences consist of short
tandem repeats that contribute to the stability
and integrity of the chromosome.
24. • Moderately repetitive DNA includes:
– variable number tandem repeats (VNTRs)
– minisatellites
– microsatellites
25. • Short interspersed elements (SINES) and long
interspersed elements (LINES) are dispersed
throughout the genome rather than tandemly
repeated, and constitute over 1/3 of the
human genome.
• These transposable elements are generated
via an RNA intermediate and are referred to as
retrotransposons.
26. • The Vast Majority of a Eukaryotic Genome
Does Not Encode Functional Genes
• Only a small portion of the eukaryotic genome
(2%–10%) constitute protein-encoding genes.
• There are also a large number of single-copy
noncoding regions, some of which are
pseudogenes.
27. FUNCTION OF CHROMATIN
“The function of the chromatin is to
carry out the genetic information
from one generation to another, by
encoding the past history and future
prospects of the cell. DNA, being the
only permanent component of
chromatin, is the sole genetic material
of eukaryotes. It never leaves the
cell, thus maintaining heredity of the
cell”
28. CONCLUSION
Chromatin is the complex combination of DNA and proteins that organizes
chromosomes which appear as many thread like coiled and elongated
structures suspended in the nucleoplasm. So the chromatin contains genetic
material instructions to direct cell function.
The first level of packing in Chromatin involves the binding of DNA to
histones into fundamental packing unit particles called nucleosomes.
The second level of packing involves packing of nucleosomes into 30 nm
thick chromatin fibre.
The highest level of packing of chromatin in the chromosome is found at
the metaphase stage of cell division.
There are two distinct types of chromatin- euchromatin
heterochromatin which differ on their staining properties.
and
In the chromatin, DNA and basic proteins called histones are present in
about equal amounts.
DNA is the permanent component of chromosomes and is the sole genetic
material of eukaryotes.