TELOMERE

Independent studies by two distinguished geneticists, Hermann J. Muller and Barbara

McClintock (although with different organisms; Muller with Drosophila and McClintock
with maize), performed in the late 1930s revealed that the ends of a chromosome are
essential for chromosome stability.
Telomeres are specialized structures which cap the ends of eukaryotic chromosomes. They
have several likely functionsmaintaining the structural integrity of a chromosome (if a
telomere is lost, the resulting chromosome end is unstable as it then becomes susceptible to
attack by exonucleases) and ensuring complete replication of the extreme ends of
chromosomes.
Studies in Tetrahymena thermophilia by Elizabeth Blackburn and Joseph Gall first
revealed that ends of the minichromosomes consist of short, simple sequence, TTGGGG,
repeated over and over. Most telomeres isolated so far are a sequence of 5 to 8 bases that is
repeated hundreds or thousands of times at each end of every chromosome. For example,
the repeat array is TTAGGG in humans and other vertebrates and TTTAGGG in Chlorella.
Most Telomeric DNA sequences have a high G content in the strand that runs 53
toward the end of the chromosome. The G-rich strand ends in a 3 single stranded
overhang. That is, the G-rich strand extends beyond the 5-end of the complementary Crich strand. The overhang contains from two to three repeat units in simple eukaryotes but
approaches 30 repeat units in humans.
The 3 overhang is vulnerable to attack by nucleases. Three different structures have been
identified that protect the 3 overhang of telomeres from degradation.
First, the G-rich DNA in telomere repeat can form complex structures.
Biochemists have discovered that four guanines can form a planar G-tetraplex,
with the four bases hydrogen-bonded to each other (Hoogsteen-type base pairs).
The 3-terminal telomeric overhang of the ciliated protozoan Oxytricha nova has
the sequence d(T4G4). The telomere end binding protein of O. nova is a
heterodimeric capping protein that binds to and protects the foregoing 3
overhang. The DNA binds in a deep cleft between the proteins and subunits,
where it adopts an irregular nonhelical conformation. The subunit contains
three so-called OB folds (OB for oligomer binding) whereas the has only one
OB fold.
Electron microscopy studies performed by Titia de Lange and coworkers in 1999
revealed that the G-rich strands 3-overhang folds back to invade the doublestranded region in the mammalian telomere to form a t-loop (telomere loop) and
a D-loop (displacement loop). Specific proteins are required to establish and
maintain the telomere structure. Although here we can focus on proteins that
protect the mammalian telomere, similar proteins are also present in other kinds
of eukaryotes. Two of the telomere proteins, TRF1 and TRF2 (telomere repeat
binding factors 1 and 2), bind to the double-stranded region of the telomere as
pre-formed homodimers. TRF1 is negative regulator of telomere length. TRF2
participates in t-loop formation and also appears to be required to cap and
protect the chromosome ends. TRF1 and TRF2 each recruit other proteins to the
telomere. A second type of DNA-binding protein, POT1 (protection of telomeres)
is probably associated with the single strand in the D-loop. This initial model
underwent modifications with more proteins being discovered. POT1 has a
binding partner called TPP1 (so named because it had been previously called

TINT1, PTOP and PIP1). Two more proteins have been discovered, TIN2 (for
TRF interacting protein) and RAP1 (for repressor activator protein 1). The
complex formed by these six proteins is known as shelterin.
The telomeres of Drosophila chromosomes are different from those of most other
organisms. Telomeres in Drosophila consist of multiple copies of two different
transposable elements, Het-A and Tart, arranged in tandem repeats. Apparently, in
Drosophila, the loss of Telomeric sequences in course of replication is balanced by the
insertion of additional copies of Het-A and Tart elements into the telomere.
The cell must maintain its telomeres. Failure to do so leads to a wide assortment of serious
problems, ranging from chromosome fusion to programmed cell death. Eukaryotic
chromosomes lose DNA from their ends as a result of replication and require a special
enzyme called telomerase to replace the lost DNA. Most human somatic cells lack
telomerase and so cannot replace the telomere DNA lost during replication. Failure to
replace this DNA is probably responsible for the fact that somatic cells go through a finite
number of divisions when studied in tissue culture, a phenomenon that is directly related
to the ageing process. Germ cells have telomerase and so can divide indefinitely. Most
cancer cells also have telomerase.
Further away from the ends of chromosomes are telomere-associated sequences,
comprising from several thousand to hundreds of thousands of base pairs. They, too,
contain repeated sequences, but the repeats are longer, more varied and more complex
than those found in telomeric sequences.