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Eukaryotic chromosome
Macromolecular organization
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Metaphase chromosome structure

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Chromatid
A pair of chromatids are clearly visible in the Metaphase chromosome Both are symmetrical They are attached at the centromere They separate at anaphase

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Chromonemata or Chromonema
Represents a chromatid in the early stages of

condensation
These are present thin filamentous structures

Each consists of a single linear DNA molecule associated

with proteins These form the gene bearing portions of the chromosomes
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Chromomeres
Bead like accumulations of chromatin
Sometimes visible during interphase Appears like a beaded string Clearly visible in the polytene chromosomes Not visible in the metaphasic chromosomes These are regions of tightly folded DNA
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Centromere and Kinetochore

Centromere lies within the primary constriction of the chromosome It is a region to which the mitotic spindle attach The regions of chromosomes flanking the centromere contain highly repetitive DNA (constitutive heterochromatin) It stains more intensely with basic dyes Centromeres are found to contain specific DNA sequences with

special proteins bound to them, forming a disc-shaped structure,

called Kinetochore
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Structure of the Kinetochore

When observed under the Electron Microscope the Kinetochore shows the following structure: Plate or cup-like disc 0.20 to 0.25nm in diameter

Situated upon primary constriction/centromere

Trilaminar structure - 10nm thick - outer proteinaceous dense layer - middle low density layer - dense inner layer tightly bound to centromere
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Outer surface of the outer layer of the kinetochore gives rise to the corona or collar of fine filaments (in

addition to microtubules)
Main function of the Kinetochore is to provide a centre of assembly for microtubules, i.e., it serves as a nucleation centre for the tubulin and hence assists their assembly into microtubules

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Based on the number of centromeres the chromosomes can be classified into the following types:
Monocentric one centromere Holocentric entire length of the chromosome acts as the centromere. Occurs in certain species of plants and animals. Most well known example is the nematode Caenorhabditis elegans Acentric no centromere Dicentric two centromeres Note: Acentric and dicentric chromosomes are abnormal chromosomes formed on exposure to ionizing radiations such as X-rays

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Telomere
Gr. Telo = far; meros = part Chromosomal ends are known as telomeres Prevent end to end fusion with other chromosomes

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Secondary constriction
Present at any point on the chromosome Constant in their position and extent Useful to identify chromosome Secondary constrictions can be distinguished from primary constriction or centromere because

chromosome bends only at the position of the

centromere at anaphase
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Nucleolar organizers
Certain secondary constrictions contain genes for 5.8s, 18s and 28s rRNA These induce formation of nucleoli In humans nucleolar organizers are located on chromosomes 13, 14, 15, 21 and 22 these chromosomes are acrocentric and have satellites
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Satellites
Round elongated or knob-like appendages
Are connected to rest of the chromosome by a thin chromatin filament Chromosomes with satellites are designated as sat chromosomes

Shape and size of satellites remain constant

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ULTRASTRUCTURE OF THE CHROMOSOME

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ULTRASTRUCTURE OF CHROMATIN
SINGLE STRANDED AND MULTISTRANDED HYPOTHESES Wide differences with respect DNA content of related species. The causes could be:

1. Lateral multiplication of chromonemata leading to multistrandedness or

2. Tandem duplication of chromonemata leading to lengthwise duplication

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Multistrandedness has been observed in plants such as

Vicia faba and salivary gland chromosomes of dipteran insects But this is not a general phenomenon hence the multistranded hypothesis was not accepted

Many evidences have supported the single stranded

nature of chromatin Each chromosome is formed from a single linear DNA molecule
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FOLDED FIBER
Folded-fiber model proposed by EJ Dupraw in 1965
Bulk of the chromosome is visualized to be composed of a tightly folded fiber which has a homogeneous diameter of 200 to 300 A The folded fiber is suggested to contain the DNA histone

helix of 30 A diameter in a supercoiled condition

Histones were proposed to be bound on the outside of the DNA coils
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NUCLEOSOME MODEL
Proposed by R.D. Kornberg, 1974
Confirmed and christened P. Oudet et.al., 1975

The DNA was found to wind around the histone particles

(the idea of histones outside the DNA coils was found to be incorrect) Most significantly and universally accepted model
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NUCLEOSOMES AND SOLENOID MODEL OF CHROMATIN

Nucleosomes are the fundamental packing unit particles of the chromatin They give the chromatin a beads-on-a-string appearance in electron micrographs

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NUCLEOSOME
Each nucleosome is a disc-shaped particle Diameter 11nm Height 5.7nm

Consists 2 copies each of nucleosome histones H2A, H2B,

H3, H4 The histone octamer forms a protein core

The DNA helix winds 13/4 times around the histone octamer
and consists of 146 bps
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2 nucleosomes are connected by a

segment of DNA called as the Linker

DNA which is 54 bp long and

contains a single H1 histone protein molecule
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Therefore, generally DNA makes 2 complete turns around the histone octamers and these 2 turns (200 bp long) are sealed off by H1 molecules

On an average nucleosomes repeat at intervals

of about 200 nucleotides/bps

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SOLENOID MODELS
SOLENOID coil of coils

Solenoid coiling of nucleosome

containing fiber 10nm fiber packing of DNA is 5-7 fold (57 times more compact than free DNA)
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30nm fiber consists of closely packed nucleosomes. Formed by folding of the nucleosome chain into a solenoid structure. Contain 6 nucleosomes per turn DNA of 30nm solenoid has 40 fold packing

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Formation of metaphase chromosome

Radial Loop model Proposed by Laemmli

Radial loops of 30nm fiber

DNA is arranged in loops anchored to a nonhistone scaffold
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KARYOTYPE & IDIOGRAM

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All members of a plant or animal species have certain constant characteristics These characteristics include: o Number of chromosomes o Their relative size o Position of the centromere o Length of the arms o Secondary constrictions and satellites
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Karyotype
Karyotype is the representation of entire
metaphase chromosomes in a cell,

arranged in descending order of size

It is the term given to the group of

characteristics that identifies a particular

set of chromosomes
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Human (male) Karyotype

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Human (female) Karyotype

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The karyotype of human somatic cells. A human somatic cell contains two sets of homologous chromosomes, which may be divided into two types:autosomes and sex chromosomes. Autosomes are further divided into seven groups: A to G. During the metaphase of cell division, each chromosome has been duplicated. Therefore, this karyotype consists of 92 chromosomes.
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Idiogram
A diagram or graphic representation of a karyotype,

showing the number, relative sizes, and morphologic

characteristics of the chromosomes of a species, individual, or cell

Gr., idios = distinctive; gramma = something written

Generally a haploid set of an organism are ordered in a series of decreasing size Sometimes an idiogram is prepared for the diploid set of chromosomes. In which the pairs of homologues are ordered in a series of decreasing size
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A karyotype of human metaphase chromosomes is obtained from their microphotographs Individual chromosomes are cut out of the

microphotographs and lined up by size

The homologous chromosomes are arranged in pairs The technique is improved by determining the centromeric index, which is the ratio of the lengths of the long and short arms of the chromosomes
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Human (male) Idiogram

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Uses of Karyotypes
Karyotypes of different species are sometimes compared and similarities in them are suggested to represent evolutionary relationship
Karyotypes also suggest primitive or advanced features of an organism They may be symmetric or asymmetric Asymmetric Karyotype - Karyotype exhibiting large differences in smallest and largest chromosomes of the set and containing fewer metacentric chromosomes e.g. Ginkgo biloba Symmetric Karyotype does not exhibit large differences between smallest and largest chromosomes of the set and contains more number of metacentric chromosomes e.g. Pinus
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An asymmetric karyotype is considered to be an advanced feature compared to the symmetric type It is suggested that in flowering plants there is a

prominent trend towards asymmetric karyotypes

This trend has been well studied in the genus Crepis of the family compositae It has been shown that increased karyotype asymmetry is associated with specialized zygomorphic flowers
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1860s and onwards

Mendel and Darwin Propose Mechanisms of Heredity

Mendel's publication of his experiments in plant

hybridization,

Darwin's provisional hypothesis of pangenesis

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1900s
German anatomist Walther Flemming recognized and explored the fibrous network within the nucleus, which he termed chromatin, or "stainable material." (Flemming had actually discovered the chromosome, although the term would be coined a few years later by Heinrich Waldeyer) Boveri and Sutton Bridges, C. B., Non-disjunction as proof of the chromosome theory of heredity Thomas Hunt Morgan Experimentally Demonstrates Chromosome

Theory - direct evidence in support of chromosome theory

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Types Of Chromatin
Depending on their staining properties, the following two types of chromatin may be distinguished in the interphase nucleus

1. Euchromatin
2. Heterochromatin

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Euchromatin
1. Euchromatin: partially condensed portions of chromosomes, that stain lightly are termed as Euchromatin It represents most of the chromatin that disperse after mitosis has completed

Euchromatin consists of structural genes (i.e., genes that

encode proteins) This is considered as genetically active chromatin since it has a role in the expression of genes DNA in euchromatin is packed into 3 to 8nm fiber
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Heterochromatin
Dark staining regions in which the chromatin remains condensed is termed as the heterochromatin
Heitz,1928 regions of the chromosome that remain condensed during interphase and early prophase and form the chromocentre Characterized by high content of repetitive DNA sequences

Very few (if any) structural genes are present

Late replicating (i.e., it is replicated after the bulk of DNA is replicated) It is not transcribed DNA is tightly packed into the 30nm fiber
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Types of Heterochromatin
Within the interphase nucleus: Condensed chromatin around the nucleolus perinucleolar heterochromatin Inside the nucleolus intranucleolar heterochromatin

Both the above mentioned heterochromatin are together called as

nucleolar heterochromatin Dense clumps of deeply staining heterochromatin in close contact with the nuclear envelop condensed peripheral heterochromatin
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Heterochromatin can be further classified into: 1. Constitutive heterochromatin 2. Facultative heterochromatin

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Constitutive heterochromatin
DNA is permanently inactive
Remains in the condensed state through out the cell cycle Occurs around the centromeres, telomeres and C-bands of chromosomes

It consists of short repeated sequences of DNA, called satellite DNA it is called so because upon ultracentrifugation, it separates from the main component of DNA
Satellite DNA may contain higher or lower G+C content than the main fraction E.g. mouse satellite DNA is a 240 bp sequence that is repeated about 1000,000 (106) times in the mouse genome, constituting 10% of the total mouse genome
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Drosophila virilis constitutive heterochromatin is present around the centromere makes up 40% of the chromosomes called pericentromeric heterochromatin In many species entire chromosome becomes heterochromatic and are called B chromosomes, satellite chromosomes or accessory chromosomes these contain very minor biological roles

Such chromosomes are found in corn, many phytoparasitic insects and salamanders
Fly Sciara large metacentric and heterochromatic chromosomes are found in the gonadal cells but are absent in somatic cells Entire Y chromosome of male Drosophila is heterochromatic although they contain six gene loci which are necessary for male fertility
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Facultative Heterochromatin
It is not permanently maintained in the condensed state Undergoes periodic dispersal and during these times it is transcriptionally active Frequently one chromosome of the pair becomes either totally or partially heterochromatic

E.g. X chromosome in the mammalian female, one of which is active and remains euchromatic, whereas the other is inactive and forms at the interphase this is called as sex chromatin and Barr body (named after its discoverer, Canadian cytologist Murray L Barr) Barr body consists of DNA which is not transcribed and is not found in males
Number of Barr bodies is always one less than the number of X chromosomes E.g. in humans XXX female has two Barr bodies and XXXX female has 8/26/2013 three Barr bodies