Lesson 12. Structure and function of the nucleus. Levels of chromatin packing.

Task 01. Nucleus structure and functions

Listen to your instructor’s explanation, read the text and draw a scheme of a cell nucleus.
Mark the nuclear envelope with inner and outer membranes, nuclear pores, nucleoplasm, chromatin
and nucleolus. Write down the main functions of the nucleus.
The nucleus contains most of the genes in the eukaryotic cell. (Some genes are located in
mitochondria and chloroplasts.) The diameter of the nucleus is about 5 μm. The nucleus is enclosed by
nuclear envelope which is a double plasma membrane. The membranes are separated by a space of 20 –
40 nm. The envelope is perforated by pore structures that are about 100 nm in diameter. At the lip of
each pore, the inner and outer membranes of the nuclear envelope are continuous. An intricate protein
structure called a pore complex lines each pore and plays an important role in the cell by regulating the
entry and exit of proteins and RNAs, as well as large complexes of macromolecules. Except at the pores,
the nuclear side of the envelope is lined by the nuclear lamina, a netlike array of protein filaments that
maintains the shape of the nucleus by mechanically supporting the nuclear envelope. The main
functions of nucleus are storage, replication and realization of the genetic information in the cell.
The nucleus contains genetic information in the form of DNA which is organized into discrete
units called chromosomes. Each chromosome contains one long DNA molecule associated with many
proteins. Some of the proteins help coil the DNA molecule of each chromosome, reducing its length and
allowing it to fit into the nucleus. The complex of DNA and proteins making up chromosomes is called
chromatin. The chromatin is surrounded by gel-like substance called nucleoplasm (or nuclear sap) which
also contains one or more nucleoli and other substances such as ions, proteins, nucleotides, etc.
Nucleolus (plural, nucleoli) is a prominent structure which the nondividing nucleus which appears
through the electron microscope as a mass of densely stained granules and fibers adjoining part of the
chromatin. Here a type of RNA called ribosomal RNA (rRNA) is synthesized from instructions in the DNA.
Also in the nucleolus, proteins imported from the cytoplasm are assembled with rRNA into large and
small subunits of ribosomes. These subunits then exit the nucleus through the nuclear pores to the
cytoplasm, where a large and a small subunit can assemble into a ribosome. Sometimes there are two or
more nucleoli; the number depends on the species and the stage in the cell’s reproductive cycle.
Nucleus structure
 Task 02. Levels of chromatin packing
Listen to your instructor explanation, read the text and draw a scheme of the levels of
chromatin packing.
Eukaryotic chromosomes each contain a single linear DNA double helix that, in humans,
averages about 1.5 × 108 nucleotide pairs. This is an enormous amount of DNA relative to a
chromosome’s condensed length. If completely stretched out, such a DNA molecule would be about 4
cm long, thousands of times the diameter of a cell nucleus. In the cell, eukaryotic DNA is precisely
combined with a large amount of protein. Together, this complex of DNA and protein, called chromatin,
fits into the nucleus through an elaborate, multilevel system of packing.
The double helix of DNA (~2nm)
(the nucleotide sequence connected by hydrogen bonds)
Nucleosomes or “beads on string” (~10 nm)
Nucleusome is the basic, bead-like unit of DNA packaging
in eukaryotes consisting of a segment of DNA wound
around a protein core. The core is composed of two
molecules of each of four types of the protein called
histone: H2A, H2B, H3 and H4. The amino end (N-
terminus) of each histone (the histone tail) extends
outward from the nucleosome.
30-nm fiber
The next level of packing results from interactions
between the histone tails of one nucleosome with the
linker DNA and nucleosomes on either side. A fifth histone,
H1, is involved at this level. These interactions cause the
extended 10-nm fiber to coil or fold, forming a chromatin
fiber roughly 30 nm in thickness, the 30-nm fiber.

Looped domains (300 nm fiber)

The 30-nm fiber, in turn, forms loops called looped
domains attached to a chromosome scaffold made of
proteins, thus making up a 300-nm fiber. The scaffold is
rich in one type of topoisomerase, and H1 molecules also
appear to be present.
Metaphase chromosome (1400 nm)
In a mitotic (dividing) chromosome, the looped domains
themselves coil and fold in a manner not yet fully
understood, further compacting all the chromatin to
produce the characteristic metaphase chromosome shown
in the micrograph above. The width of one chromosome is
1400 nm. Particular genes always end up located at the
same places in metaphase chromosomes, indicating that
the packing steps are highly specific and precise.

Even during the interphase (the period between nuclear divisions), some chromosomal regions (such as
centromeres, telomers, etc.) exist in a highly condensed state similar to that seen in a dividing
(metaphase) chromosome. This type of interphase chromatin, visible as irregular clumps with light
microscopes is called heterchromatin, to distinguish it from the less compacted euchromatin (“true
chromatin”). Because of its compaction, heterochromatic DNA is largely inaccessible to the machinery in
the cell responsible for transcribing the genetic information coded in the DNA, a crucial early step in
gene expression. In contrast, the looser packing of euchromatin makes its DNA accessible to this
machinery, so the genes present in euchromatin can be transcribed.
 Task 03. The structure and function of chromosomes
Read the text below and make a sketch of duplicated chromosomes showing their main parts:
centromere (or primary constriction), kinetochore, short arm (p), long arm (q), telomere.
Chromosome of eukaryotes is thread-like structure that made up of a macromolecule of nuclear
DNA associated with protein, found in the nuclei of most living cells. Chromosomes play an important
role in storing, processing and replication of cell genetic information. Chromosomes are highly dispersed
and therefore they are not visible under the light microscope in non-dividing (interphase) cells. During
mitosis chromosomes replicate and condensed through coiling (so become visible in a light microscope)
forming chromosomes consisting of two sister chromatids joined at the centromere (or primary
constriction). Centromere is a region containing specific DNA sequences (heterochromatin) which is
responsible for the accurate segregation of the replicated chromosome during cell division. The part of
the centromere to which spindle microtubules attach during cell division is called kinetochore.
Centromere divides chromosome into two arms. If the arm differ in length, then the short arm is
designated "p" (for petite), and the long arm is designated "q" (because it follows the letter "p").
Telomeres are long stretches on non-coding DNA (heterochromatin) at the ends of chromosomes.
Telomeres provide terminal stability to the chromosome and ensure its survival. If chromosome breaks,
the broken ends do not contain telomeres, so they can stick with each other. Telomeres also assist in
the pairing of homologous chromosomes and crossing over.

Figure. Chromosome structure. Duplicated chromosomes before nucleus division.

Some chromosomes have secondary constrictions which can be located at any place of their
arms. Secondary constrictions are constant in their position and extent and therefore prove useful to
identify a particular chromosome in a set. Secondary constrictions may arise because the rRNA genes
are transcribed very actively and thus interfering with chromosomal condensation. A secondary
constriction can separate a distal region of a chromosome arm forming nob-like round or elongated
structures which called satellites. Satellites keep connected to the rest of the chromosome by a thin
filament. Shape and size of satellites remains constant.
 Task 03. Chromosome types
Read the text and draw the main types of eukaryotic chromosomes (metacentric,
submetacentric, acrocentric and telocentric).
Chromosomes can be divided into four types based on the centromere position. In metacentric
(met-uh-CEN-trick) chromosomes, the centromere lies near the centre of the chromosome so that the
arms of such chromosome are roughly of equal length. Submetacentric (SUB-met-uh-CEN-trick)
chromosomes have a centromere that is off-centre, so that one chromosome arm is longer than the
other. Acrocentric chromosomes have their centromere have very near one end. Therefore, the p-arm
of the chromosome becomes really short making it very difficult to observe. Telocentric chromosomes
have a centromere that is located at the very tip of one end. Thus, telocentric chromosomes have a 'rod'
shaped appearance. Humans do not possess telocentric chromosomes.

Figure. Types of chromosome based on the location of centromere.