| ODUCTION TO TRANSPOSONS | | | | over evolutionary time. These Interspersed Repeats |
| (Jumping Genes) | | | | block gene conversion thereby catalyzing the |
| Author: Waseem Ashfaq | | | | formation of new genes. Transposons are therefore |
| Transposons are pieces of DNA | | | | an evolutionary device promoting creation of new |
| that can “jump” into novel positions in the | | | | genes and protecting novel gene sequences from |
| genome. There are numerous different types of | | | | being overwritten by similar gene sequences through |
| transposon which differ from each other in many | | | | gene conversion. |
| ways. They have been exploited very extensively in | | | | There are three distinct types: |
| molecular biology research. They can insert | | | | · Class II Transposons consisting only of DNA |
| themselves into DNA without requiring any similar | | | | that moves directly from place to place. |
| sequence to be present in themselves and their | | | | · Class III Transposons; also known as Miniature |
| “target”. This makes them powerful | | | | Inverted-repeats Transposable Elements or MITEs. |
| mutagens, since their insertion into a gene will | | | | · Retrotransposons (Class I) that |
| generally prevent the gene from producing a | | | | -first transcribe the DNA into RNA |
| functional protein. As mutagens, their effect is not | | | | and then |
| always completely random, as some do have | | | | -use reverse transcriptase to make a |
| preferences for certain sequences or types of | | | | DNA copy of the RNA to insert in a new location. |
| chromatin, and others will transpose preferentially to | | | | Many transposons move by a "cut |
| adjacent regions rather than randomly throughout the | | | | and paste" process: the transposon is cut out of its |
| genome. Generally, a minimum of two things are | | | | location (like command/control-X on your computer) |
| required for transposition. | | | | and inserted into a new location (command |
| The first is a gene which | | | | control-V). This process requires an enzyme — a |
| encodes transposase enzyme and which can | | | | transposase — that is encoded within some of |
| recognize the ends of the transposon, excise the | | | | these transposons. Transposase binds to: |
| transposon (or a copy of it) from its starting point, | | | | · both ends of the transposon, which consist of |
| cut DNA randomly or semi-randomly elsewhere, and | | | | inverted repeats; that is, identical sequences reading |
| catalyze the movement of the transposon from its | | | | in opposite directions. |
| starting point to its new target. The transposase can | | | | · a sequence of DNA that makes up the target |
| be encoded either by the transposon itself (in which | | | | site. Some transposases require a specific sequence |
| case the transposon is often referred to as being | | | | as their target site; other can insert the transposon |
| “autonomous”) or elsewhere – on | | | | anywhere in the genome. |
| another transposon, for example, or on a plasmid. | | | | After the transposon is ligated to the host |
| Transposons which rely on a transposase which they | | | | DNA, the gaps are filled in by Watson-Crick base |
| do not themselves carry are called | | | | pairing. This creates identical direct repeats at each |
| “non-autonomous”. They occur naturally, and | | | | end of the transposon. Often transposons lose their |
| any autonomous transposon can be converted to a | | | | gene for transposase; but as long as somewhere in |
| non-autonomous one by removing the transposase | | | | the cell there is a transposon that can synthesize the |
| gene from it. The second requirement for | | | | enzyme, their inverted repeats are recognized and |
| transposition are the sequences at the ends of the | | | | they, too, can be moved to a new location. |
| transposon, which are recognized by the transposase. | | | | Retrotransposons move by a "copy |
| This minimal requirement for an autonomous | | | | and paste" mechanism but in contrast to the |
| transposon is shown below. There are many variants | | | | transposons described above, the copy is made of |
| on this theme! Transposons use many different | | | | RNA, not DNA. The RNA copies are then |
| mechanisms to jump around the genome, but they all | | | | transcribed back into DNA — using a reverse |
| boil down to two basic types: replicative and | | | | transcriptase — and these are inserted into new |
| nonreplicative. Replicative transposons copy | | | | locations in the genome. Many retrotransposons |
| themselves when they transpose, and leave one | | | | have long terminal repeats (LTRs) at their ends that |
| copy at the original site while inserting the second | | | | may contain over 1000 base pairs in each. Like DNA |
| copy elsewhere. Non-replicative transposons are | | | | transposons, retrotransposons generate direct |
| excised from their starting position and inserted | | | | repeats at their new sites of insertion. In fact, it is |
| elsewhere in the genome. | | | | the presence of these direct repeats that often is |
| In replicative transposition a), the | | | | the clue that the intervening stretch of DNA arrived |
| transposon makes a copy of itself which is inserted | | | | there by retrotransposition. 42% of the entire human |
| randomly in the target DNA. In non-replicative | | | | genome consists of retrotransposons. |
| transposition b), the transposon excises from its | | | | Retroviruses were first identified |
| original position (usually leaving a small change in the | | | | 80 years ago as agents involved in the onset of |
| sequence at the excision point) and reinserts at | | | | cancer. More recently the AIDS epidemic has been |
| random in the target DNA. | | | | shown to be due to the HIV retrovirus. In the early |
| They may be: | | | | 1970s it was discovered that retroviruses had the |
| · cause mutations | | | | ability to replicate their RNA genomes via conversion |
| · Increase (or decrease) the amount of | | | | into DNA which became stably integrated in the DNA |
| DNA in the genome. | | | | of the host cell. It is only comparatively recently that |
| These mobile segments of DNA | | | | retroviruses have been recognized as particularly |
| are sometimes called "jumping genes". They were | | | | specialized forms of eukaryotic transposons. In |
| discovered by Barbara McClintock early in her career, | | | | effect they are transposons which move via RNA |
| for which she was awarded a Nobel prize in 1983. | | | | intermediates that usually can leave the host cells and |
| The chromosomal basis of heredity was already well | | | | infect other cells. The integrated DNA form (provirus) |
| established by the time McClintock began her | | | | of the retrovirus bears a marked similarity to a |
| graduate training in the Botany Department at Cornell | | | | transposon. |
| University. Her experiments laid the groundwork for a | | | | The transposition cycle of |
| serie of cytogenetic discoveries by the Cornell maize | | | | retroviruses has other similarities to prokaryotic |
| genetics group between 1929 and 1935. McClintock | | | | transposons, which suggest a distant familial |
| developed a method for using broken chromosomes | | | | relationship between these two types of transposon. |
| to generate new mutations. Among the progeny of | | | | Crucial intermediates in retrovirus transposition are |
| plants that had received a broken chromosome from | | | | extrachromosomal DNA molecules. These are |
| each parent, she observed unstable mutations at an | | | | generated by copying the RNA of the virus particle |
| unexpectedly high frequency, as well as a unique | | | | into DNA by a retrovirus-encoded polymerase called |
| mutation that defined a regular site of chromosome | | | | reverse transcriptase. The extra chromosomal linear |
| breakage. These observations so intrigued her that | | | | DNA is the direct precursor of the integrated element |
| she began an intensive investigation of the | | | | and the insertion mechanism bears a strong similarity |
| chromosome-breaking locus. Within several years she | | | | to "cut and paste" transposition. |
| had learned enough to reach the conclusion, published | | | | One family of transposons in the |
| in 1948, that the chromosome-breaking locus did | | | | fruit fly Drosophila melanogaster are called P |
| something unknown for any genetic locus: it moved | | | | elements. They seem to have first appeared in the |
| from one chromosomal location to another, a | | | | species only in the middle of the twentieth century. |
| phenomenon she called transposition. | | | | Within 50 years, they have spread through every |
| Transposons are discrete | | | | population of the species. Transposons are also a |
| segments of DNA capable of moving through the | | | | widely used tool for mutagenesis of most |
| genome of their host via an RNA intermediate in the | | | | experimentally tractable organisms. |
| case of class I retrotransposon or via a | | | | In bacteria, transposons can |
| "cut-and-paste" mechanism for class II DNA | | | | jump from chromosomal DNA to plasmid DNA and |
| transposons. Since transposons take advantage of | | | | back, allowing for the transfer and permanent |
| their host's cellular machinery to proliferate in the | | | | addition of genes such as those encoding antibiotic |
| genome and enter new hosts, transposable elements | | | | resistance (multi-antibiotic resistant bacterial strains |
| can be viewed as parasitic or "selfish DNA". However, | | | | can be generated in this way). Bacterial transposons |
| transposons may have been beneficial for their hosts | | | | of this type belong to the Tn family. When the |
| as genome evolution drivers, thus providing an | | | | transposable elements lack additional genes, they are |
| example of molecular mutualism. Some transposable | | | | known as insertion sequences. The most common |
| elements contain heat-shock like promoters and their | | | | form of transposon in humans is the Alu sequence. |
| rate of transposition increases if the cell is subjected | | | | The Alu sequence is approximately 300 bases long |
| to stress, thus increasing the mutation rate under | | | | and can be found between 300,000 and a million |
| these conditions, which might be beneficial to the cell. | | | | times in the human genome. |
| The evolution of transposons and | | | | Mariner-like elements are another |
| their effect on genome evolution is currently a | | | | prominent class of transposons found in multiple |
| dynamic field of study. Transposons are found in all | | | | species including humans. The mariner transposon was |
| major branches of life. The study of transposable | | | | first discovered by Jacobson and Hartl in drosophila1. |
| genetic elements and transposition became the | | | | This type-II trasposable element is known for its |
| central theme of her genetic experiments from the | | | | uncanny ability to be transmitted horizontally in many |
| mid 1940s until the end of her active research career. | | | | species. There are an estimated 14 thousand copies |
| This was incredulous at the time, DNA was believed | | | | of mariner in the human genome composing of 2.6 |
| to be stable and invariable. They may or may not | | | | million base pairs. These characteristics of the mariner |
| have originated in the last universal common | | | | transposon have inspired the science fiction novel |
| ancestor, or arisen independently multiple times, or | | | | titled, "The Mariner Project." |
| perhaps arisen once and then spread to other | | | | References: |
| kingdoms by horizontal gene transfer. While | | | | 1. McClintock, B. (June 1950). "The origin and |
| transposons may confer some benefits on their | | | | behavior of mutable loci in maize". Proc Natl Acad Sci |
| hosts, they are generally considered to be selfish | | | | U S A. 36 (6): 344–55. |
| DNA parasites that live within the genome of cellular | | | | 2. Rubin GM, Spradling AC (October 1982). |
| organisms. In this way, they are similar to viruses. | | | | "Genetic transformation of Drosophila with |
| Viruses and transposons also share features in their | | | | transposable element vectors". Science 218 (4570): |
| genome structure and biochemical abilities, leading to | | | | 348–53. |
| speculation that they share a common ancestor. | | | | 3. Wilson MH, Coates CJ, George AL (January |
| Since excessive transposon activity | | | | 2007). "PiggyBac transposon-mediated gene transfer |
| can destroy a genome, many organisms seem to | | | | in human cells". Mol. Ther. 15 (1): 139–45. |
| have developed mechanisms to reduce transposition | | | | 4. Mandal, P.K. & Kazazian, H.H., Jr. SnapShot: |
| to a manageable level. Bacteria may undergo high | | | | Vertebrate transposons. Cell 135, 192-192 e1 (2008). |
| rates of gene deletion as part of a mechanism to | | | | 5. Kidwell, M.G. (2005). "Transposable elements.". |
| remove transposons and viruses from their genomes | | | | in (ed. T.R. Gregory). The Evolution of the Genome. |
| while eukaryotic organisms may have developed the | | | | San Diego: Elsevier. pp. 165–221. ISBN |
| RNA interference (RNAi) mechanism as a way of | | | | 0-12-301463-8. |
| reducing transposon activity. In the nematode | | | | 6. Craig NL, Craigie R, Gellert M, and Lambowitz |
| Caenorhabditis elegans, some genes required for | | | | AM (ed.) (2002). Mobile DNA II. Washington, DC: ASM |
| RNAi also reduce transposon activity. | | | | Press. ISBN 978-1555812096. |
| Interspersed Repeats within genomes | | | | 7. Lewin B (2000). Genes VII. Oxford University |
| are created by transposition events accumulating | | | | Press.. ISBN 978-0198792765. |