NUCLEOLUS AND IT’S

ORGANIZATION
EXPLORING THE NUCLEUS
INTERIOR…………..
 CONTENTS
► What is a nucleolus?????
► The milestones of early nucleolus research ….
► Features of nucleolus
► Morphology of nucleolus
► Evolution of tripartite organization
► Nucleolar organization and dynamic
► Nucleolus in mitosis
► The plurifunctional nucleolus
► The Nucleolus and Human diseases
► Conclusion
 What is a nucleolus?????
► A prominent sub-nuclear
structure that is not bound
by a membrane and resides
within the nuclear matrix.

► The site of the biogenesis of

ribosomal RNA (rRNA) and
assembly of ribosomes.

► Called as “Ribosome
Factories”

Source: Molecular Expressions:

micro.magnet.fsu.edu/.../nucleolus
 The milestones of early
nucleolus research ….
► 1835-Discovery of the nucleolus (termed "Keimfleck" or
"macula germinatova") in germinal vesicles
► 1839-Introduction of the term "nucleolus" (small nucleus or
nucleus of the nucleus).
► 1893-Observation that the nucleoli gradually disappear
during prophase.
► 1934-Demonstration of special chromosomal regions called
secondary chromosomal constrictions or nucleolus-organizing
regions (NORs).
► 1952-First isolation of nucleoli.
► 1962-After a pulse of radioactive precursor, the label is
initially found in 45S RNA and then transferred from 45S to
28S RNA.
► 1969-The spreading technique made it possible to visualize
the transcription process. Each transcription unit looks like a
"Christmas tree".
 Features of nucleolus
► Dark stained body in the nucleus

► Formed by the nucleolar organizing regions (NOR) of DNA

► A dynamic structure: Nucleoli are not static structures. They

disassemble during mitosis and reform in early G1 phase .

► Size and number vary according to the requirement of cell.

► A site for ribosome biogenesis.

 Morphology of nucleolus
Nucleoli have got tripartite
organization composed of
three morphologically
distinct regions :-

► Fibrillar Center (FC)

► Dense Fibrillar Center or

Dense Fibrillar Component
(DFC)

► Granular region or
Granular Component (GR)

Fig: The FC(asterisks), the DFC(arrow) and

GC(G) Bar: 0.5 μm
(source: review-springerlink springer-
Verlag 200510.1007/s00418-005-0046-4)
 Fibrillar Center (FC)
► lightly stained when
observed by EM
► composed of "fibrils" (±
50Ǻ)
► contains DNA that is not
actively being transcribed
► presence of pol I
► multiple FC in one nucleolus
► their number doubles in G2
compared to G1
► accounts for only 1-2
percent of the total volume
of the nucleolus
 Dense Fibrillar Center (DFC)

► Surround the FC’s

► Composed of "densely
packed fibrils" (30-50 Ǻ)
► Contains RNA molecules
being transcribed
► Occupies a large fraction of
the nucleolus, ± 17 percent.

Fig: The dense fibrillar component (arrow)

and the granular component (G). Bar:
0.3 μm
(source: review-springerlink springer-
Verlag 200510.1007/s00418-005-0046-4)
 Granular region (GR)
► Region encompassing both
the FC and the DFC
► Consisting of granules 150-
200 Ǻ
► Contains maturing ribosomal
precursor particles
► Granule rich region due to the
presence of RNP particles
► With a fraction of about 75
percent, it occupies the
largest fraction of the total
nucleolus volume
► It separates nucleolus and
nucleoplasm

Fig: The dense fibrillar component (arrow)

and the granular component (G). Bar:
0.3 μm (source: review-springerlink
springer-Verlag 200510.1007/s00418-005-0046-
 Evolution of tripartite
organization

► The "tripartite’ organization" (FC, DFC, GC) of the nucleolus is

commonly accepted.
► Proposed that this particular organization is only observed in
higher eukaryotes
► It evolved from a bipartite organization with the transition
from anamniotes to amniotes.
► Reflecting the substantial increase in the rDNA intergenic
region, an original fibrillar component would have separated
into the FC and the DFC (Thiry and Lafontaine 2005).
Other components of Nucleolus
► A substantial (additional) component of the nucleolus is
chromatin, which penetrates the organelle from the
surrounding nucleoplasm.

► One last structure identified within the nucleolus is nucleolar

vacuole. There are multiple nucleolar vacuoles in the
nucleolus, but it remains unclear whether or not they serve
some functional or structural purpose.

► Several sub-nuclear compartments are often associated with

nucleoli including, the perinucleolar compartment (PNC); the
Cajal and Sam68 bodies; and paraspeckles.
 Nucleolar organization and
dynamics
► Transcription of ribosomal DNA (rDNA) by RNA polymerase I
occurs either in the fibrillar centers (FCs) or at the boundary
between the FC and the dense fibrillar component (DFC)
region

► Pre-ribosomal RNA transcripts are spliced and modified by

small nucleolar ribonucleoproteins (snoRNPs) in the DFC. This
is revealed by the presence of constituting proteins, for
example fibrillarin, nucleolin.

► Final maturation of the pre-ribosomal ribonucleoprotein and

assembly with ribosomal proteins occurs mostly in the
granular component (GC) region. Protein B23 and NOP52 in
GC provide evidence for it.
 Nucleolus in mitosis
 Early prophase- cyclin B1–
CDK1 levels increase, the
transcription machinery
usually remains attached to
active NORs during mitosis,
some RNA polymerase I
subunits leave the fibrillar
centre (FC).

 Late prophase- early and

late processing factors and
partially processed pre-
ribosomal RNAs (pre-rRNAs)
leave the nucleolus at the
same time.

Metaphase- the majority of

processing components are
associated with the surface Review: The multifunctional nucleolus
of chromosomes as a www.nature.com/.../v8/n7/images/nrm2184-f6.jpg
Anaphase- cytoplasmic processing components become
packaged in nucleolar-derived foci (NDF), whereas the other
components remain around the condensed chromosomes. In late
anaphase, cyclin B1–CDK1 levels decrease.
Early telophase- the number of NDF decreases and prenucleolar
bodies (PNBs) are formed on the surface of each chromosome.
The PR breaks down (indicated by an interrupted line) and
processing components are taken up by PNBs. Nucleoli start to re-
form around NORs of acrocentric chromosomes.
Late telophase- the nuclear envelope is re-formed and early (1)
and late (2) processing components relocate in an ordered
manner to the dense fibrillar component (DFC) and granular
component (GC), respectively. CDK1, cyclin-dependent kinase-1;
rRNA, ribosomal RNA.
 The plurifunctional nucleolus
 Major role as ribosome factories

 Virus infection control: Many viruses target nucleolar functions

as part of their infectious strategy. The Rev protein of human
immunodeficiency virus (HIV) and the Rex protein of human T-
lymphotrophic virus (HTLV-I) depend on an interaction with B23,
an endogenous cellular nucleolar protein .

 Maturation of non-nucleolar RNAs or RNPs: e.g. RNase P RNA

 Regulation of telomerase function

 Tumor suppressor and oncogene activities

 cell stress sensing and signaling

 Role in cell senescence (aging)

 Biosyntheses of signal recognition particle RNA and

telomerase RNA involve a nucleolar stage

 Involved in processing of U6 RNA, one of the spliceosomal

small nuclear RNAs.

 Regulation of cell cycle.

 The Nucleolus and Human
diseases
 The nucleolus has been linked to multiple forms of human
disease.
 Multiple genetic disorders have been mapped to human genes
that encode nuclear proteins that are known to associate with
nucleoli under specific conditions, including Werner syndrome
and Bloom syndrome, more recently, Rothmund–Thomson
syndrome.
 Humoral hypercalcaemia of malignancy is a common
complication of lung and certain other cancers. PTHrP was
first discovered as the hypercalcaemia factor that is produced
 by solid tumor.
Diseases It localizes
linked to the nucleus
with aberrant ribosomeand biogenesis.
the nucleolus in
certain tissues,
Mutations such
in genes as skin,
other thancartilage and
those that bone.DNA
encode
helicases also link the nucleolus to disease pathogenesis, as
shown by Diamond–Blackfan anaemia.
 Conclusion
► Nucleolus is a dynamic entity.
► Structure correlates with function.
► Has role in processes other than ribosome
biogenesis also.
► Still a lot to find out more about structure and
function.
► New imaging techniques will provide deeper
understanding
 Bibliography
 Hernandez-Verdun, D. 2006a. [http://www.springerlink.com/content/75n545v0g3186830
[http://www.springerlink.com/content/75n545v0g3186830 Nucleolus: From structure to
dynamics. Histochem Cell Biol 125: 127-137. Retrieved July 8, 2008.
 Hernandez-Verdun, D. 2006b. The nucleolus: A model for the organization of nuclear functions.
functions. Histochem Cell Biol
126: 135-148. Retrieved July 8, 2008.
 Khadzhiolov, A. A. 1985. The Nucleolus and Ribosome Biogenesis. Wien: Springer-Verlag. ISBN 3211817905.
3211817905.
 Olson, M. O. J. 2004. The Nucleolus. Georgetown, TX: Landes Bioscience/ Eurekah.Com. New York: Kluwer
Academic/Plenum Publishers. ISBN 0306478730.
0306478730.
 Olson, M. O. J., and M. Dundr. 2005. The moving parts of the nucleolus.
nucleolus. Histochem Cell Biol 123: 203-216. Retrieved
July 8, 2008.
► McKeown PC,PC, Shaw PJ.
PJ.
► Chromatin: linking structure and function in the nucleolus.
► chromosoma, 2008 Oct 17 ,springerlink review.

► EMBO Rep. 2006 September; 7(9): 870–873. Published online 2006 August 18. doi:
10.1038/sj.embor.7400786.PMCID: PMC1559673Copyright © 2006, European Molecular Biology Organization
► Olson, M.O., M. Dundr, and A. Szebeni. "The Nucleolus: An Old Factory with Unexpected Capabilities." Trends in Cell
Biology (2000) 10: 189-196.
► Websites
► www.google.com
► www.springerlink.com
► www.ncbi.nlm.nih.gov