LECTURE- 10

PART- I CYTOGENETICS

HOMEOBOX GENES THE

MOLECULAR ARCHITECTS

Homeobox genes Architects of body plan

How does a single fertilized cell develops into a complex
organism like a fly, a mouse, or a human being?
Von Baer in the early 19th century observed that all vertebrates look

very similar in their early stages of embryonic development.

Geoffroy Saint-Hilaire remarked that all animals have the same body

plan. As the main nerve cord is in the front part of insects and in the
back part of vertebrates, he hypothesized that vertebrates are
essentially upside-down invertebrates!

Homeobox genes Architects of body plan

Homeotic genes (homeo = alike) or homeobox genes
are similar in structure and function in all animals.

They are an important class of regulatory genes.

They serve as molecular architects and direct the

building of body segments according to definite detailed
plans.

Homeobox genes What are they?

The genes are tandemly repeated to form a homeobox
gene family.

They code for proteins that bind to DNA; have a homeobox

sequence- conserved DNA motif of about 180 base pairs.

Their proteins contain a homeodomain (DNA binding

domain- 60 amino acids long), and a variable domain.

Homeobox genes produce DNA-binding

proteins

The homeodomain
contains a helix-turnhelix DNA-binding motif
characteristic of many
DNA-binding proteins.

Role Homeobox genes

Homeobox genes and their encoded homeodomain proteins
play important roles in the developmental processes of
multicellular organisms.

They play crucial roles from the earliest steps in

embryogenesis such as cell differentiation and

organization within segments:
- e.g. the differentiation of neurons in the nematode
(Caenorhabditis elegans).

Mechanism of function of Homeobox genes

The homeobox genes can be thought of as genetic
switches or master control genes that turn different
programs of cellular differentiation on or off.

Homeobox proteins are transcription factors that

upregulate the transcription of other genes by binding to
their upstream elements.

Mechanism of function of Homeobox genes

Order of development of multicellular organisms

Polarity- formation of the axis by which embryo differentiates.

Before fertilization an egg has a gradient of proteins that help

to establish its polarity (anterior- head and posterior- tail).
After fertilization Maternal Effect genes reinforce polarity and
also establish the dorsal (back) and ventral (belly) orientation.
Segmentation occurs driven by Gap genes, Pair rule Genes
and Segmentation polarity genes.
Finally the homeotic genes are switched on.

Order of development in Drosophila

Gap genes
Establishes domains of distinct identity along the anteriorposterior axis.
The gap domains are multiple segments in width.
Pair-rule and segment polarity genes
These genes merely subdivide the embryo into parasegments
and organize short-range pattern within a parasegment
Homeotic genes
Homeotic genes assign distinct identities to different
parasegments, by initiating preprogrammed differentiation of
each segment.

Regulation of Drosophila homeotic gene complex

Homeotic genes interact with a variety of information from
the gap, pair-rule, and segment polarity genes.
Mutations in pair-rule genes have also been shown to affect
homeotic gene expression

Cross-regulation between homeotic genes: homeotic genes

also regulate each other.
Cross-regulatory interactions are important in defining the domains of
expression.
More posterior acting genes function as negative regulators of their
more anterior neighbors.

The homeotic genes in Drosophila

The activities of a number of homeotic genes are required to establish
the identity of parasegments in the trunk region (posterior head, thorax,
and abdomen) of the embryo.

These genes are clustered in two major groups called the:

Antennapedia complex (ANT-C)- responsible for segmental

identity in the head and anterior thorax
AND
Bithorax complex (BX-C)- responsible for segmental identity in
the posterior thorax and abdomen.

The homeotic genes in Drosophila

The five Drosophila genes that belong to the ANT-C are:
- Labial (lab)
- Proboscipedia (pb)
- Deformed (Dfd)
- Sex combs reduced (Scr)

-Antennapedia (Antp)

The homeotic genes in Drosophila

The three Drosophila genes that belong to the BX-C are:
- Ultrabithorax (Ubx)
- Abdominal-A (Abd-A)
- Abdominal-B (Abd-B)
Within the ANT-C and BX-C, the order of the genes on the
chromosome is the same as the order of segments that they

affect along the embryonic axis.

This is referred to as the "colinearity principle".

Colinearity principle

The homeobox gene family comprises a cluster of genes that encodes a specific body
part. The posterior ones regulate the anterior ones.

ANT-C homeotic gene mutation of Drosophila

ANTP controls development of middle segment of the thorax
(mesothorax) of Drosophila.
Mesothorax produces a pair of legs that are distinct from the forelegs
and hindlegs.
ANTP encodes a homeodomain regulatory protein that is expressed in
the mesothorax during embryo development.
But a dominant ANTP mutation caused by a chromosome inversion,
brings ANTP protein-coding sequence under the control of regulatory
DNA that mediates gene expression in the head tissues.

ANT-C homeotic gene mutation of Drosophila

This mutation results in legs developing instead of antennae in the
head region.
Hence mutations in an ANT-C gene result in gross organizational
changes.

Bithorax complex (BX-C)

UBX (ultrabithorax) encodes a homeodomain regulatory
protein that controls the development of the 3rd thoracic
segment (metathorax).

UBX represses genes responsible for the development of the

2nd thoracic segment (mesothorax).
UBX regulates ANTP; represses ANTP expression in the
metathorax and restricts its expression to mesothorax only.

Bithorax complex (BX-C)

UBX mutants cannot regulate ANTP expression.

This results in ANTP also being expressed in the metathorax

and transforming it into a second mesothorax.

Mesothorax has a pair of legs and wings; while metathorax

has a pair of legs and halteres (balance flies during flight)

UBX mutants have 2 pairs of wings!!!

Bithorax complex (BX-C)

A mutation called CBX (contrabithorax) causes UBX to be
expressed in the mesothorax.

CBX mutants look like wingless ants.

Vertebrate Hox genes

In vertebrates HOX genes are found in gene clusters on
the chromosomes. Mammals have 4 Hox clusters
(Hoxa, Hoxb, Hoxc, Hoxd), organized into thirteen
homology groups.

Mice and humans contain 38 Hox genes arranged in

these 4 clusters.

Vertebrate Hox genes- human

Vertebrate Hox genes- mouse

Vertebrate Hox genes- mouse and human- same

The Vertebrate Hox Gene Complex

Only about 38 genes out of a total of about 30,000 control
most of the development, architecture, and appearance of
the body plan of complex mammalian species.

The mouse Hox complex is exactly similar to the human

Hox complex.

The Vertebrate Hox Gene Complex

The organization of the genes in each cluster reflects its
anterior-posterior expression in the body plan (spatial
colinearity).

Unlike in Drosophila, vertebrate Hox genes are also

temporally colinear in addition to being spatially colinear.

Homeotic genes are expressed within segmented and

unsegmented structures within the body plan.

Mutations in human Hox genes

Human Hox gene mutations
have shown to affect limb
development.
A mutation in the human
Hoxd13 gene results in
polydactyly.

Mutation in Hox gene can also

result in an extra rib.

The Hox genes and the evolution of the eye

Swiss biologist, Walter Gehring and his team, found that the
Hox gene responsible for induction of the Drosophila eye is
virtually identical to the one that induces the mouse eye.

This Hox gene switches on eye formation in the myriad of

creatures that see. Hence, it appears that all eyes, no
matter how differently constructed they appear now, had a
common evolutionary origin.

Homeobox genes have a wide phylogenetic

distribution
Homeobox genes have a wide phylogenetic distributionfound in baker's yeast, plants, and all animal phyla that
have been examined so far.

The incredible conservation of genes across species

suggests that the homeobox gene clusters and their role
in development are of ancient origin.

Homeobox genes have a wide phylogenetic

distribution
All invertebrates and vertebrates came from a bilateral
ancestor with 7 Hox genes that lived 600 million years ago.
In insects, a gene near the right end of the cluster was
duplicated.

In vertebrates, the entire Hox cluster was duplicated:

3 times in mammals
up to 8 times in some types of fish.

The duplicate genes were then free to take on new functions,

often leading to more-complex body structures.

1 billion years ago

600 million years ago

Homeobox genes have a wide phylogenetic

distribution

The differences in homeobox gene clusters that are

seen between species only occur with respect to the

number of clusters as well as to the number of genes
involved in each cluster.

Homeobox genes have a wide phylogenetic

distribution
Hence, Flies and people are just variations on a theme of how to
build a body plan that was laid down in some worm-like creature
in the Cambrian period."

If a gene plays a central role in development in the fly, it is worth

determining whether it plays a similar role in another organism.

Comparing genes by sequence homology have proved extremely

successful in elucidating the role and identity of several human

homeobox genes.