CENTROMERES AND TELOMERES

It has been known for several decades that the telomeres (from the Greek terms telos and
meros,
meaning “end” and “part,” respectively), or ends of eukaryotic chromosomes, have unique
properties.
Hermann J. Muller, who introduced the term telomere in 1938, demonstrated that Drosophila
chromosomes without natural ends—produced by breaking chromosomes with X rays—were
not transmitted to progeny. In a classical study of maize chromosomes, Barbara McClintock
demonstrated that new ends of broken chromosomes are sticky and tend to fuse with each
other.
In contrast, the natural ends of normal (unbroken) chromosomes are stable and show no
tendency to fuse with other broken or native ends. McClintock’s results indicated that
telomeres must have special structures different from the ends produced by breakage of
chromosomes.
Another reason for postulating that telomeres have unique structures is that the known
mechanisms of replication of linear DNA molecules do not permit duplication of both strands
of DNA at the ends of the molecules. Thus, telomeres must have unique structures that
facilitate their replication, or there must be some special replication enzyme that resolves this
enigma. Whatever their structure, telomeres must provide at least three important functions.
They must:
(1) Prevent deoxyribonucleases from degrading the ends of the linear DNA molecules,
(2) Prevent fusion of the ends with other DNA molecules
(3) Facilitate replication of the ends of the linear DNA molecules without loss of material.
The telomeres of eukaryotic chromosomes have unique structures that include short
nucleotide sequences present as tandem repeats. Although the sequences vary somewhat in
different species, the basic repeat unit has the pattern 5’ T1–4A0–1G1–8-3’ in all but a few
species. For example, the repeat sequence in humans and other vertebrates is TTAGGG that
of the protozoan Tetrahymena thermophila is TTGGGG, and that of the plant Arabidopsis
thaliana is TTTAGGG. In most species, additional repetitive DNA sequences are present
adjacent to telomeres; these are referred to as telomere-associated sequences.
In vertebrates, the TTAGGG repeat is highly conserved; it has been identified in more than
100 species, including mammals, birds, reptiles, amphibians, and fishes. The number of
copies of this basic repeat unit in telomeres varies from species to species, from chromosome
to chromosome within a species, and even on the same chromosome in different cell types. In
normal (noncancerous) human somatic cells, telomeres usually contain 500 to 3000
TTAGGG repeats and gradually shorten with age. In contrast, the telomeres of germ-line
cells and cancer cells do not shorten with age.
The telomeres of a few species are not composed of short tandem repeats of the type
described earlier. In D. melanogaster, for example, telomeres are composed of two
specialized DNA sequences that can move from one location in the genome to other
locations. Because of their mobility, such sequences are called transposable genetic elements.
Most telomeres terminate with a G-rich single-stranded region of the DNA strand with the 3’
end (a so-called 3’ overhang). These overhangs are short (12 to 16 bases) in ciliates such as
Tetrahymena, but they are quite long (50 to 500 bases) in humans. The guanine-rich repeat
sequences of telomeres have the ability to form hydrogen-bonded structures distinct from
those produced by Watson–Crick base pairing in DNA. Oligonucleotides that contain tandem
telomere repeat sequences form these special structures in solution, but whether they exist in
vivo remains unknown.
The telomeres of humans and a few other species have been shown to form structures called
t-loops, in which the single strand at the 3_ terminus invades an upstream telomeric repeat
(TTAGGG in mammals) and pairs with the complementary strand, displacing the equivalent
strand. The DNA in these t-loops is protected from degradation and/or modification by DNA
repair processes by a telomere-specific protein complex called shelterin. Shelterin is
composed of six different proteins, three of which bind specifically to telomere repeat
sequences. TRF1 and TRF2 bind to double-stranded repeat sequences, and POT1 (Protection
Of Telomeres 1) binds to single-stranded repeat sequences. Subunits TIN2 and TPP1 tether
POT1 to DNA-bound TRF1 and TRF2, and the TRF2-associated protein Rap1 helps regulate
telomere length. Shelterin is present in sufficient quantities in most cells to coat all the single-
and double-stranded telomere repeat sequences in the chromosome complement.
To date, t-loops have been identified in the telomeres of vertebrates, the ciliate Oxytricha
fallax, the protozoan Trypanosoma brucei, and the plant Pisum sativum (peas). Thus, they are
probably important components of the telomeres of most species.
 Electron micrograph of a human metaphase chromosome
from which the histones have been removed. A huge pool of
DNA surrounds a central “scaffold” composed of non-
histone chromosomal proteins. Note that the scaffold has
roughly the same shape as the metaphase chromosome
prior to removal of the histones. Also note the absence of
ends of DNA molecules in the halo of DNA surrounding the
scaffold.

TELOMERE LENGTH AND AGING IN HUMANS

Unlike germ-line cells, most human somatic cells lack or have very low levels of telomerase
activity. When human somatic cells are grown in culture, they divide only a limited number
of times (usually only 20 to 70 cell generations) before senescence and death occur. When
telomere lengths are measured in various somatic cell cultures, a correlation is observed
between telomere length and the number of cell divisions preceding senescence and death.
Cells with longer telomeres survive longer—go through more cell divisions— than cells with
shorter telomeres. As would be expected in the absence of telomerase activity, telomere
length decreases as the age of the cell culture increases. Occasionally, somatic cells are
observed to acquire the ability to proliferate in culture indefinitely, and these immortal cells
have been shown to contain telomerase activity, unlike their progenitors. Since the one
common feature of all cancers is uncontrolled cell division or immortality, scientists have
proposed that one way to combat human cancers would be to inhibit the telomerase activity
in cancer cells.
Further evidence of a relationship between telomere length and aging in humans has come
from studies of individuals with disorders called progerias, inherited diseases characterized
by premature aging. In the most severe form of progeria, Hutchinson–Gilford syndrome,
senescence—wrinkles, baldness, and other symptoms of aging—begins immediately after
birth, and death usually occurs in the teens. This syndrome is caused by a dominant mutation
in the gene encoding lamin A, a protein involved in the control of the shape of nuclei in cells.
Why this mutation leads to premature aging is unknown. In a less severe form of progeria,
Werner syndrome, senescence begins in the teenage years, with death usually occurring in the
40s. Werner syndrome is caused by a recessive mutation in the WRN gene, which encodes a
protein involved in DNA repair processes. Again, we still do not understand how the loss of
this protein leads to premature aging. However, the somatic cells of individuals with both
forms of progeria have short telomeres and exhibit decreased proliferative capacity when
grown in culture, which is consistent with the hypothesis that decreasing telomere length
contributes to the aging process.
At present, the relationship between telomere length and cell senescence is entirely
correlative. There is no direct evidence indicating that telomere shortening causes aging.
Nevertheless, the correlation is striking, and the hypothesis that telomere shortening
contributes to the aging process in humans warrants further study.

John Tacket, 15, of Bay City, Michigan, speaks about

his illness, progeria, during a news conference called
in Washington, April 16, 2003, to announce the
discovery of the gene that causes this rare, fatal
genetic condition, characterized by the appearance of
accelerated aging. To Tacket’s right is Dr. Francis S.
Collins, director of the National Institutes of Health.