Molecular Mechanism of Cell

Differentiation
Two main mechanisms can be envisaged
1. Those in which genes are differentially
activated(transcriptional, post transcriptional
or translational levels)
2. Those in which the genes themselves are
altered (by amplification, DNA
rearrangement & methylation)
Control at the level of transcription
Polytene chromosomes- in which transcription
can be directly visualised in the form of puffs.
Specialised cells have distinct patterns of
puffing & therefore of transcription.
 Transcriptional control has been clearly proven
also for those genes that code for abundant
specialised proteins such as globin, ovalbumin &
silk fibroin.
To make hybridization probes fro these genes the
mRNA is copied into DNA with reverse
transcriptase & then the cDNA is cloned into
plasmids.
With these hybridisation probes, transcripts from
the corresponding gene can be detected only in
those cells that produce the protein product.
Enormous amounts of protein can be produced
from the transcriptional induction of a single
protein-coding gene because stable mRNA
molecules can be translated many times.
 Chromatin structure is one possible level at
which transcription may be controlled.

Genes in condensed heterochromatin are

inactive, frequently in a very stable form.

Nuclease digestion studies suggest that the

chromatin of active genes is more susceptible
to nuclease attack than that of inactive genes.
 DNA methylation has also been a likely
candidate to control transcription.
Eukaryotic DNA contains some 5-methylcytosine
which is always next to guanosine.
The enzyme that methylates DNA prefers
templates that already have a methyl group on
the other strand, because it provides a stable
signal on the DNA, which can be transmitted to
daughter cells after DNA replication.
In many cases genes have been shown to be
more methylated than active ones.
 The post transcriptional processing of the
transcripts is also of considerable regulatory
importance.
of the transcripts made in cells are not
polyadenylated & are discarded.
 Translational control
Cells have mechanisms by which some mRNAs can
be translated in a given cell type but not in
others.
Embryologists suspect the existence of masked
mRNAs in eggs whose translation is activated
only after fertilization.
Sea urchin eggs increase 10-fold their protein
synthesis after fertilization and this is due to
stored maternal mRNA because the increase in
protein synthesis is not inhibited by blocking new
transcription with actinimycin D.
These mRNAs are thought to be bound to proteins
that prevent their translation.
 The clearest eg of translational regulation is
provided by the heat shock response of Xenopus
oocytes.
When the frog oocytes are exposed to high
temperature, the synthesis of a major heat shock
protein of 70,000 daltons is induced while the
synthesis of the other endogenous proteins is
repressed.
Here synthesis is accomplished by a translational
mechanism ie by activation of the translation of
stored (but masked) heat shock mRNA.
This is known because oocytes can still respond in
the same way after manual removal of the cell
nucleus & because heat shock mRNA can be
detected biochemically in oocytes that do not
synthesize the corresponding proteins at the normal
temperature.
 Gene Amplification
One way of obtaining differential gene activity would
be to increase the no. of copies of a specific gene.
There are 3 circumstances in which gene
amplification is known to occur:

1. Ribosomal DNA amplification observed in the

oocytes of amphibians & insects
Xenopus oocytes selectively replicate their rDNA genes; a
mature oocyte has 2 million copies of them (compared
to 900 for a diploid somatic nucleus) and 1000 nucleoli
inorder to produce the vast no of ribosomes (1012 )
contained in a single egg.
2. DNA replication observed in certain puffs of the
dipteran Rhynchosciaria salivary gland polytene
chromosome.

In these DNA puffs, in addition to an increased RNA

synthesis as in all other puffs there is incorporation
of thymidine.
DNA amplification can be visualized by an increase in
the amount of DNA in the particular polytene band
when the puff collapses after the phase of active
RNA synthesis is over.
3. Egg shell proteins of Drosophila
The egg chorion is formed by the follicle cells during a
period of 5 hours.
It has been found that the follicle cells replicate more of a
90 kb segment of DNA containing the chorion protein
genes, so that finally it is 16 times more abundant than
the rest of the DNA.
Although it is clear that the selective gene amplification
does indeed occur in all known cases the specially
amplified DNA is not passed on to future cell
generations.
The larval polytene salivary glands die at metamorphosis,
the follicle cells die when the egg is laid & the amplified
oocyte rDNA is not inherited by the frog embryo.
 Transposable genes in yeast &
trypanosomes

There are cases in which DNA rearrangements

cause stable changes in patterns of gene
expression.
Eg: mating type switch in yeast
 Bakers yeast generally propagates as haploid
cells, but at a low frequency 2 cells may fuse &
produce diploids that are able to produce spores
if life becomes hard.
The cells can fuse only if they are of opposite sex
or mating type. The mating types are a and .
When haploid strains of one mating type are
cultured, some cells are transformed into cells of
the opposite sex with a certain frequency.
If the latter are cultured again, they can switch
sex again.
Furthermore these changes are stable & persist
over several cell generations.
 Genetic information for the a or mating
types is stored in silent places in yeast
chromosome. These storage sites are called
silent cassettes & their expression is activated
only when these genes are transposed into
the exposed position on the MAT mating
type of locus.
The silent genes are copied before their
transposition & therefore a copy remains at
their original position.