Homeotic Genes on Evolution Development: How does a Fly get a Leg as its Antennae?

Samantha Ongchuan Panther Creek High School 6770 McCrimmon Parkway Cary, NC 27519-1838

Abstract The study of Homeotic or Hox genes is the leading forefront in 21st century evolution and development, popularly coined as evo-devo. Advances in genomic sequencing have enabled studies in homeotic genes through manipulation of hox cluster in Drosophila melanogaster. Hox genes are transcription factors that control the expression of genes along the antero-posterior axis. Diversity today from an ancestral organism could be attributed to hox genes duplication throughout evolution under environmental pressure. Correlations between an organisms number of hox clusters and the complexity and/or diversity of that particular organism shed light how gene duplication and evolution may have shaped evolution of morphological structures. Furthermore, changes in hox expression and protein function have also played an important role in animal evolution.

For the most part, the genome seems to be a welter of various genes scattered about randomly, with no order present in their arrangement on a chromosomethe order only becomes apparent in their expression through the process of development. The Hox genes, in contrast, seem like an island of comprehensible structure. (Myers, 2008) Homeotic, or Hox genes, are genes that determine the placement of parts along the anterior-posterior axis. They are important in laying down the body plan for all animals and are the basic blocks for evolution of development. Hox genes were discovered in the Drosophila melanogaster by Ernst Hafen, Michael Levine, and William McGinnis in 1983, at the Universityof Basel, Switzerland. Edward Lewis from Pasadena, California was also a leading investigator of hox genes. He discovered the bithorax complex in Drosophila melanogaster and won the Nobel Peace prize for medicine in 1995 with Christiane Volhard, and Eric Wieschaus for his work on hox genes. Though hox genes were the leading forefront of the first half of the twenty-first century in evolution development, it was not until genomic sequencing was more readily available, that researchers fully grasped the significance of the hox genes; it connected countless organisms through its homology from the first ancestral organism. Hox genes was a milestone for evolution development, popularly coined as evo-devo which focuses on comparing different development processes of organisms to determine ancestral relations between them. Evolution development stems from the earlier studies of phylogeny (study of evolution relatedness) and ontology (study of origin). Even Aristotle had thoughts about evolution development. He had two theories on embryonic development; one was that the embryo came preformed and just increased in size over time and the other was epigenesist. Epigensis is similar to what we think of embryonic development today: new

structures develop progressively. Another aspect of evolution development is Haeckels biogenetic law, also known as the theory of recapitulation, which states that embryonic development in the young in any given species reflects the evolutionary history in that species (Wolpert, 2009).Though Haeckel may have overemphasized the similarity between phylogeny and ontogeny with his embryo drawings, the concept that embryo research can provide evidence of the morphological changes of an organisms past is not so far fetched. Hox genes are one of the best evidences that twenty-first century has given us pointing that all animals come from a common ancestor: Hox genes were discovered through homeotic mutations which is when the cells predetermined fate is switched during the embryonic development. A classic example of this was found in the Drosophila melanogaster (fruit fly), in which a leg formed where the antennae should have been formed. This involves the Hox family HOM-C which is organized into the Antennapedia complex and the bithorax complex. The Antennapedia controls foot and leg placement while the bithorax complex controls abdominal and neck placement. Mutations are induced. The bithorax complex (200 kilobases) and Antennepedia complex (300 kilobases) have both been mapped out and compared. (Marx) The presence of the ftz gene and homeotic genes indicate that the two complexes may have originated from the same ancestral complex. Many believe that the antennepedia complex was the anterior homolog of the bithorax complex and was created through gene duplication. Moreover, similar Hox clusters were found in different species such as a frogs, mice, and even humans. This indicates that they originated from an ancestral hox gene and evolve through tandem and genome duplications. Hox gene study is important because it gives a glimpse at how genes evolved by comparing their function between divergent organisms.

Hox genes are homeobox containing genes; the homeobox, a 180 bp sequence, encode a that encode 60 amino acid motif called the homeodomain The homeodomain is the part of the hox protein that bind to the DNA double helix. . Homeobox protein recognizes and bind to the following DNA sequences TAAT with T being the most important for binding. In the genome, Hox genes are organized into clusters. Vertebrates normally have four distinct clusters that house up to 13 Hox genes. Each cluster is located on a different chromosome and the clusters are the results from the genome duplication that preceded the evolution of the vertebrates. Hox genes are organized in paralogous groups which contain hox genes that occupy the same location on different clusters. There are up to 13 paralogous group. Examples of Hox genes are HOX-A10, HOX-A13, HOXB1, etc. For example, HOX-A10 functions in fertility and embryo development. Also, the number of Hox genes in each organism varies. Humans have 39 Hox genes organized in 4 clusters while fruit flies only have eight Hox genes organized in one cluster. In interesting characteristic about hox genes is that their genomic organizations (location on the cluster) mimic their expression during embryonic development. Hox genes located on the 3 end of the cluster will be expressed early and more anteriorly while genes located at the 5 end will express later and more posteriorly. In addition, posterior genes are the more dominant gene in relation to anterior genes. The placement of hox genes is not random; they have a very distinct organization in the genome in which the order of the genes within the cluster corresponds to some aspect of gene expression. Ed Lewis noticed that, in the bithorax complex of D. melanogaster, the homeotic genes were strategically staggered in accordance to their expression along the posterior-anterior axis. This specific placing is called spatial colinearity. Another type of hox gene placement is

temporal colinearity. Temporal colinearity states that the 3/Anterior side of the DNA strand will develop first and the 5/Posterior side will form later in embryonic development. Hox genes start to be expressed in mesoderm cells at the early stage of gastrulation. The anterior proteins will develop first and the posterior proteins next in accordance to temporal collinearity. As the posterior qualities start to define, hox expression are best seen in the mesoderm and nueral tube after somite formation and neurulation (Wolpert, 2009). HOX proteins also need cofactors such as extradenticle and homothorax to select and regulate their target genes. Nuclear localization happens when exd and homothorax bind together and allow the HOX proteins to bind to DNA. Specifically, Extradenticle and DNA bind cooperatively with the Hox protein; however, homothorax is needed so that extradenticle can perform nuclear localization which enables the complex to bind to specific DNAtarget sequence. (Ryoo, 1999) Though Hox genes are not found in sponges, protozoa, and plants, they are prevalent in all bilaterian (symmetric sides) and aquatic organisms. Todays clusters of hox genes are believed to have originated from one hox gene from the first organism through spontaneous gene duplication. Gene duplication and diversification created three major bilaterian families: deuterostomes (humans, dogs, sea urchins), ecdysozoans (insects, spiders, crustaceans), and lophotrochozoans (mollusks, segmented worms). The oldest known bilateral ancestor was believed to have eight hox genes including the Evx class. The Evx gene was also found in the fruit fly, Drosophila melanogaster. However this ancestral Evx class was not conserved in most animals. Instead, the Evx gene takes on a new role in serving axial functions; it was called Evx/Hx to distinguish from its ancestral counterpart. The

change in function of Hox genes indicate that Hox genes can be altered over time attributing to the diversity of animals today from its original ancestor. The sequencing of animal genomes has constantly reminded the scientific community of the ability of the homeotic gene to preserve its clustered organization. By comparing the number and type of hox clusters of one organism to that of another, evolutionists can deduct the ancestry and history of that particular organism. There have been some contradicting correlations, but interesting correlations nonetheless. For example, teleoset fish have seven hox clusters due to a lineage specific whole genome duplication they have gone through in the past while tetrapods have maintained the 4 clusters architecture. On the other hand, the low numbers of hox clusters in nematodes (round worms) prove the simple structure of the animal compared to its more complex relatives. In addition, losses of certain hox genes prove a difference in the structure of the animal; in Sacculina carcini the loss of abdominal segments is correlated with the loss of abdominal A-Hox gene. The study of hox genes is not only interesting because of its distinct effect in the morphology of developing embryos, but it is also very important because it provides a plethora of evidence that change in hox expression and protein functions have instigated morphological changes in animal evolution. Hox genes enable evolutionists to place ancient organisms on their respective places in the phylogenic tree with more conviction. Hox gene studies can even give us a glimpse of the future. Theoretically, with enough environmental pressure, the mammalian species can gain an extra hox cluster to promote diversity and thus survival. Who knows what we might become in the next ten million years?

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Special Thanks to Joe Moss and John Luke Scemama