Unit II Microbes structure and Multiplication

Organization and Structure of Microorganisms

Phylogenetic relationships amongst different cell types

 Based on ribosomal RNA sequence comparsions (16S, 23S)

 3 basic groups or domains established (domains are a higher order than

kingdoms, ie are superkingdoms)

 The 3 domain = Bacteria, Archaea and Eucarya

 3 domains are related to each other; progenote = hypothetical ancient

universal ancestor of all cells.

 Natural relationships amongst cells established (phylogeny)

Microbes have different shapes and is of advantage

 Cell wall establishes the shape of a microbial cell but environmenta

conditions can change it

 Shapes include:
o Spheres called cocci (greek = berry) can divide once in one axis to
produce diplococci (Neisseria gonnorrhoeae, N. meningitidis), or more
than once to produce a chain (Streptococcus pyogenes), divides
regularly in two planes at right angles to produce a regular cuboidal
packet of cells (xxx) or in two planes at different angles to produce a
cluster of cells (Staphyloccus aureus)
o Cylinders called rods or bacilli (Latin bacillus = walking stick)
o Spiral or spirilli (Greek spirillum = little coil)

 Shape offers an advantage to the cell:

o Cocci: more ressistant to drying than rods
o Rods: More surface area  & easily takes in dilute nutrients from the
environment
o Spiral: Corkscrew motion & therefore less ressistant to movement
o Square: Assists in dealing with extreme salinities

 
Microbes are small but this feature is crucial

 Nutrients and wastes are transported in and out the cell via the cytoplasmic
membrane.

 The rate of transport determines the metabolic rates and therefore the
growth rates of microbial cells

 The smaller the size, the larger the surface area of the cytoplasmic
membrane to volume and therefore the faster is it's potential growth rate.
This can be seen as follows:

 
radius (r) of cell A  =  radius (r) of cell B =
 
1um 2um
Surface area (SA) of cell =
  12.6um2 50.3um2
4pir2
Volume (V) of cell = 4/3pir3  4.2um3 33.5um3
Ratio of SA to V  3  1.5

Diagramatic representation of cells

Cell walls are external structures that shape and protect cells

1. Bacterial Cell Walls:

 All the members of domain Bacteria, with the exception of the genera
Mycoplasma, Ureaplasma, Spiroplasma, and Anaeroplasma contain cell
walls

 Cell walls are chemically peptidoglycans ie peptides (short amino acids

chains) and glycans (sugars); peptidoglycans  are a.k.a. mureins,
mucopeptide.
o Glycans:
 are modified sugars viz, N-acetyl muramic acid (NAM or M) &
N-acetly glucose amine (NAG or G)
 M and G are linked to each other by a beta 1, 4 glycosidic bond
&  alternate to form the wall backbone.
 Lysozyme (an enzyme produced by organisms that consume
bacteria, and normal body secretions such as tears, saliva, &
egg white = protect against would-be pathogenic bacteria)
digests beta 1,4 glycosidic bonds.
 Lysozyme lyses growing or non growing cells but cell wall-less
microbes are not affected
 High osmotic pressure in high solute concentrations prevents
lysis of Gram +ve & Gram -ve cells when treated with 
lysozyme:
 spheroplasts  = part of cell wall  removed (Gram
-ve)
 protoplasts = complete removal of cell wall
(easier for Gram +ve)

o Peptides:
 Short peptides (4 amino acids, tetrapeptides) attached to M.
 Some of the amino acids are only found in cell walls & not in
other cellular proteins (D- amino acids, eg D-alanine &
diaminopimelic acid, DAP)
 Tetrapeptides chains are cross linked (interlinked) by a
peptide bridge (the carboxyl group of one tetrapeptide with an
amino group of an adjacent (direct interbridge) or a different
tetrapeptide chain (indirect interbridge).
 Transpeptidase enzyme builds peptide bridges in actively
dividing cells; penicillin binds to it stoping cell wall synthesis.
Autolysins restructure and reshape cell walls by breaking
specific bonds in the peptidoglycan in actively growing cells.
Cell wall synthesis stops but cell degrading enzymes still
function resulting in weakened cell walss and ultimately death.

 Glycans and peptides therefore forms a single, large and strong cross-linked
molecule in a form of a multilayered sheet, (sacculus, Latin = little sac) that
surrounds the entire bacterial cell.
Differences Between Gram-positive And Gram-negative Bacterial Cell Walls
 Gram-positive wall  Gram-negative wall
 Peptidoglycan  Thick layer  Thin layer
 All contain
Peptidoglycan tetrapeptide  Most contain lysine
diaminopimelate
Peptidoglycan cross  Generally via
 Direct bonding 
linkage pentapeptide
Teichoic acid  Present  Absent
Teichuronic acid  Present   Absent
Lipoproteins  Absent  Present
LPS  Absent   Present
Outer Membrane  Absent  Present
Periplasmic Space  Absent  Present
 

Virus structure
At the simplest level, the function of the outer shells (CAPSID) of a virus
particle is to protect the fragile nucleic acid genome from:

 Physical damage - Shearing by mechanical forces.

 Chemical damage- UV irradiation (from sunlight) leading to chemical
modification.
 Enzymatic damage - Nucleases derived from dead or leaky cells or
deliberately secreted by vertebrates as defence against infection.

The protein subunits in a virus capsid are multiply redundant, i.e. present in
many copies per particle. Damage to one or more subunits may render that
particular subunit non-functional, but does not destroy the infectivity of the
whole particle.

Furthermore, the outer surface of the virus is responsible for recognition of the
host cell. Initially, this takes the form of binding of a specific virus-attachment
protein to a cellular receptor molecule. However, the capsid also has a role to
play in initiating infection by delivering the genome from its protective shell in a
form in which it can interact with the host cell.

To form an infectious particle, a virus must overcome two fundamental problems:

1. To assemble the particle utilizing only the information available from the
components which make up the particle itself (capsid + genome).
2. Virus particles form regular geometric shapes, even though the proteins from
which they are made are irregularly shaped.
How do these simple organisms solve these difficulties? The information to answer these
problem lie in the rules of symmetry.

In 1957, Fraenkel-Conrat & Williams showed that when mixtures of purified tobacco
mosaic virus (TMV) RNA & coat protein were incubated together, virus particles
formed. The discovery that virus particles could form spontaneously from purified
subunits without any extraneous information indicated that the particle was in the free
energy minimum state & was therefore the favoured structure of the components. This
stability is an important feature of the virus particle.

Although some viruses are very fragile & are essentially unable to survive outside the
protected host cell environment, many are able to persist for long periods, in some cases
for years.

Helical capsid

Tobacco mosaic virus (TMV) is representative of one of the two major structural classes
seen in viruses of all types, those with helical symmetry. The simplest way to arrange
multiple, identical protein subunits is to use rotational symmetry & to arrange the
irregularly shaped proteins around the circumference of a circle to form a disc.
Multiple discs can then be stacked on top of one another to form a cylinder, with the virus
genome coated by the protein shell or contained in the hollow centre of the cylinder.

Closer examination of the TMV particle by X-ray crystallography reveals that the
structure of the capsid actually consists of a helix rather than a pile of stacked disks.

A helix can be defined mathematically by two parameters:

 amplitude (diameter)   &
 pitch (the distance covered by each complete turn of the helix)

Helices are rather simple structures formed by stacking repeated components with a
constant relationship (amplitude & pitch) to one another - note that if this simple
constraint is broken a spiral forms rather than a helix - unsuitable for containing a virus
genome.
Icosahedral (isometric) capsids:

An alternative way of building a virus capsid is to arrange protein subunits in the form of
a hollow quasi-spherical structure, enclosing the genome within. The criteria for
arranging subunits on the surface of a solid are more complex than those for building a
helix.

Francis Crick & James Watson (1956), were the first to suggest that virus capsids are
composed of numerous identical protein sub-units arranged either in helical or cubic
(=icosahedral) symmetry.

In order to construct a capsid from repeated subunits, a virus must 'know the rules' which
dictate how these are arranged. For an icosahedron, the rules are based on the rotational
symmetry of the solid, which is known as 2-3-5 symmetry:

viruses

Many viruses have devised strategies to exit the infected cell without its total destruction.
This presents a difficulty in that all living cells are covered by a membrane composed of
a lipid bilayer. The viability of the cell depends on the integrity of this membrane.
Viruses leaving the cell must therefore allow this membrane to remain intact & this is
achieved by extrusion (budding) of the particle through the membrane, during which
process the particle becomes coated in a lipid envelope derived from the host cell
membrane & with a similar composition:
The structure underlying the envelope may be based on helical or icosahedral symmetry
& may be formed before or as the virus leaves the cell. In the majority of cases,
enveloped viruses use cellular membranes as sites allowing them to direct assembly. The
formation of the particle inside the cell, maturation & release are in many cases a
continuous process.
The site of assembly varies for different viruses. Not all enveloped viruses bud from the
cell surface membrane, many viruses use cytoplasmic membranes such as the golgi
complex, others such as herpesviruses which replicate in the nucleus may utilize the
nuclear membrane. In these cases, the virus is usually extruded into some form of
vacuole, in which it is transported to the cell surface & subsequently released.

Envelope Proteins:

If the virus particle became covered in a smooth, unbroken lipid bilayer, this would be its
undoing. Such a coating is effectively inert, & though effective as a protective layer
preventing desiccation of or enzymatic damage to the particle, would not permit
recognition of receptor molecules on the host cell. Therefore, viruses modify their lipid
envelopes by the synthesis of several classes of proteins which are associated in one of
three ways with the envelope:

 Matrix Proteins: These are internal virion proteins whose function is effectively
to link the internal nucleocapsid assembly
 Glycoproteins: These are transmembrane proteins, anchored to the membrane by
a hydrophobic domain & can be subdivided into two types, by their function:
o External Glycoproteins - Anchored in the envelope by a single
transmembrane domain. Most of the structure of the protein is on the
outside of the membrane, with a relatively short internal tail. Often
individual monomers associate to form the 'spikes' visible on the surface
of many enveloped viruses in the electron microscope. Such proteins are
the major antigens of enveloped viruses.
o Transport Channels - This class of proteins contains multiple
hydrophobic transmembrane domains, forming a protein-lined channel
through the envelope, which enables the virus to alter the permeability of
the membrane, e.g. ion-channels.

Complex Virus Structures

The majority of viruses can be fitted into one of the three structural classes outlined
above, i.e. those with helical symmetry, icosahedral symmetry or enveloped viruses
based on either of these two.

However, there are many viruses whose structure is more complex. In these cases,
although the general principles of symmetry already described are often used to build part
of the virus shell (this term being appropriate here since such viruses often consist of
several layers of protein & lipid), the larger & more complex viruses cannot be simply
defined by a mathematical equation as can a simple helix or icosahedron.

Because of the complexity of some of these viruses, they have defied attempts to
determine detailed atomic structures using the techniques described earlier.

Algal structure
 I. General characteristics [For a detailed review of the Plant Kingdom, see the site
maintained by Cardillo & Samuels]
A. Eukaryotic; placed in Kingdom Protista (also frequently termed
Protoctista)
B. Mostly photosynthetic
1. Photosynthetic pigments- four different kinds of chlorophyll
2. accessory pigments- a variety, including blue, red, brown, golden
C. Require moist environments (lack a waxy cuticle found in terrestrial
plants)
D. May be microscopic and float in surface waters (phytoplankton) or
macroscopic and live attached to rocky coasts (seaweeds)
1. Size ranges from size of bacteria (0.5 um) to over 50 m long (1 um
= 1/25,000th inch; 1 m = 39 inches)
E. Lack vascular (conducting) tissues- no true roots, stems, or leaves
F. Modes of reproduction
1. Sexual and asexual
-Have single-celled gametangia (reproductive organs)- no
multicellular reproductive organs
- Life history has 1, 2, or 3 stages (in contrast, plants have 2 stages,
gametophyte and sporophyte)
View of their phylogenetic relationships
II. Representative algae
A. Red algae (Division Rhodophyta)
1. Evolution: Red algae are some of the oldest eukaryotic organisms
on the planet. Fossils of red algae have been found that are over 2
billion years old.
2. Habitat: There are 4000 different species of red algae.
a. They are very abundant in tropical and warm waters,
although many are found in cooler waters.
b. Red algae are typically found in marine waters attached to
rocks or other plants in the calmer, deeper waters beyond
the tidal zone.
c. Some red algae are reef builders in tropical seas, as
important or more important than coral animals.
d. The red algae act as habitat and food for some animals.
3. Structure: Their size and complexity vary from thin films growing
on rocks to complex filaments or membranes growing to heights
approaching 1 meter.
a. Their accessory pigments called phycobilins mask the
chlorophyll a and give them their red color. Due to these
specialized pigments, red algae are often able to
photosynthesize in deeper water than other algae.
b. Red algae do not have flagella at any stage of their life
cycle.
 4. Commercial: Chondrus crispus (Irish moss) is a small filamentous
red algae found on rocky ledges in the subtidal zone of the coastal
waters.
a. The Irish started using small quantities of Irish moss boiled
with milk to produce a jelly dessert that the French later
called "blanc mange." The mixture of a polysaccharide
from the walls of the algae with the proteins in milk
produces carrageenan.
b. This thick solution or gel is used in ice cream, whipped
cream, fruit syrups, chocolate milk, bread, and macaroni. It
is also used in tooth paste, pharmaceutical jellies, and many
kinds of lotions.
c. In this country, Irish moss is commercially harvested in
Maine.
d. The agar used to grow bacteria and other media, such as the
bread mold in the fungi experiment, is derived from red
algae.
e. Some red algae are eaten by humans.
B. Diatoms (Golden-brown algae; Division Bacillariophyta)
1. Largest group of algae but many of its species still undescribed.
2. Evolution- A relatively recent group; diatoms did not exist in the
age of the dinosaurs.
3. Habitat: cool marine oceans
a. Very important in food chains- especially in cooler, marine
waters as phytoplankton
4. Structure: mostly unicellular
a. Silica in cell walls (tiny glass houses); cell walls fit like a
Petri dish [slow to load but worth it!]
5. Reproduction- asexual for several generations, then sexual (to
restore size)
Tired of studying real diatoms? Try these!
C. Kelps (brown algae; Division Phaeophyta)
1. Evolution- Closely related to diatoms and also a young group, but
very different in appearance.
2. Habitat: rocky coasts in temperate zones or open seas (cold water
algae)
3. Structure: multicellular only. Some attain great size- 180 feet and
grow 2 feet per day.
4. Examples: shoreline: Laminaria; open ocean: Sargassum
D. Dinoflagellates (Division Pyrrhophyta or Dinophyta)
1. Evolution
2. Habitat: Especially important in food chains in warm, tropical
oceans
3. Structure: Mainly unicellular.
a. Green and colorless forms (some phagotrophic, some
parasitic- e.g. fish kills on the Pokomoke river by Pfiesteria
 b. Biflagellate
c. Nucleus unusual- chromosomes always visible
d. Some bioluminescent forms- light up when water is
disturbed
e. Reproduction commonly asexual
E. Green algae (Division Chlorophyta)
1. Next to the golden-brown diatoms, the green algae are the second
largest group of algae.
2. They are also the most diverse of the algae, with at least 7000
species.
3. Evolution- almost as old as red algae
4. Habitat: They are found mostly in fresh waters and on land. Most
species float in rivers, lakes, reservoirs, and creeks.
a. They can also live on rocks, soil, and tree bark.
b. A few species, such as sea lettuce (Ulva), live in the salt
water along the coast. Large, thin sheets of sea lettuce often
totally obscure the muddy bottom in sheltered bay and
estuary habitats.
5. Structure: Green algae are organisms with a variety of body forms
including single cells, filaments, colonies, and thalli (singular -
thallus, multicellular forms that have a leaf-like shape).
a. The higher terrestrial plants arose from a green algal
ancestor. They possess the same photosynthetic pigments
(chlorophyll a and b) and some green algae have stiff cell
walls composed of cellulose, as do plants.
6. Commercial:
a. Green algae are an important source of oxygen and food for
aquatic organisms.
b. Some are consumed as food by humans.
7. Examples - fresh water: Volvox, Spirogyra; marine: sea lettuce
(Ulva)
III. Significance to humans
A. Beneficial algae
1. Base of the aquatic food chain- especially important are the
diatoms and dinoflagellates
a. Seaweeds are not only food, but shelter for aquatic organisms-
especially important are the kelps, which form underwater forests;
Sargasso Sea community
b. Some red algae are reef builders in tropical seas; as important or
more important than coral animals
2. Other uses of importance and their history (e.g., fertilizer, fodder,
etc.) and algal farming
B. Harmful algae- excess growth causes:
1. Clogging of waterways, streams, and filters- when water is
polluted with nutrients such as fertilizer or sewage
a. A bad taste to water when present in large numbers
 b. Toxicity to animals (paralytic shellfish poisoning, red tides-
both caused by dinoflagellates)
C. Commercial uses of algae
1. Algin- thickening agent in ice cream, marshmallows- from brown
algae; carrageenan- in foods, puddings, laxatives, toothpaste- from
red algae
2. Iodine- from brown algae
3. Agar- from red algae
4. Food- especially reds and browns- important in East Asia
5. Diatomaceous earth- used for filtering, insulating, and
soundproofing

Fungal structure

Nutrition

Fungi (kingdom Fungi) are heterotrophs. They cannot manufacture their

own food as photosynthetic organisms can.

Most species of fungi are saprotrophic; they decompose dead matter.

Many are parasitic; they obtain nutrients from living organisms.

Fungi are the principle decomposers in every ecosystem. They can break
down most organic compounds including lignin, a compound that is a
major component of of wood and is very difficult to break down or digest.

Some species are parasites and others are mutualistic.

They have extracellular digestion by secreting enzymes into environment

and absorbing the nutrients produced.

Fungi store their food as glycogen (like animals). Plants and green algae
store their food as starch.

Structure

Yeasts are single-celled but most fungal species are multicellular.

Multicellular fungi are composed of filaments called hyphae (singular:

hypha).

Hyphae may contain internal crosswalls, called septa, that divide the
hyphae into separate cells. Coenocytic hyphae lack septa. The septa of
many species have pores, allowing cytoplasm to flow freely from one cell
to the next. Cytoplasmic movement within the hypha provides a means to
transport of materials. 
 The hyphae may be branched. A dense mass of hyphae is called a
mycelium.

Fungi have cell walls (like plants) but the cell walls are composed of
chitin, which is what arthropod (insects, crayfish, etc.) exoskeletons are
composed of. The cell walls of plants and some protists are composed of
cellulose.

The hyphae of some symbiotic fungi become specialized for penetrating

the cells of the host. These hyphae are called haustoria.

Most fungi do not have flagella in any phase of their life cycle. They move
toward food by growing toward it.

Reproduction

Fungi are categorized into phyla (divisions) based on the type of structures
produced during sexual reproduction. 

Some fungal species have not been classified into phyla based on
evolutionary relationships because they do not have a sexual phase or
because details regarding their sexual reproduction are unknown. They are
placed in a separate group called Deuteromycota. When details concerning
their evolutionary relationships become available, they are reclassified
into one of the other phyla.

In general, the life cycle involves the fusion of hyphae from two
individuals, forming a mycelium that contains haploid nuclei of both
individuals. The fusion of hyphae is called plasmogamy. The fused
hyphae containing haploid nuclei from two individuals is heterokaryotic.
In some cases, plasmogamy results in cells with one nucleus from each
individual. This condition is called dikaryotic. Eventually, two nuclei that
originated from different individuals fuse to form a diploid zygote.
Meiosis then produces either four haploid nuclei or four haploid cells.

The diagram below shows the generalized life cycle of fungi.

 Spores are reproductive cells that are dispersed by wind. They are capable
of germinating and producing a new mycelium.

Ecology: Some Important Symbiotic Relationships

Lichens

Lichens are structures made up of two different species: 1) a fungus and 2)

either a cyanobacterium or a green algae.

The photosynthetic cells are contained within the middle layer.

The photosynthetic cells provide photosynthesis for the lichen. It was

thought that the relationship was mutualistic because the fungus prevented
the algal cells from desiccation. Recent evidence indicates that the
photosynthetic cells may grow faster when separated from the fungus.
Perhaps the fungus is parasitizing the photosynthetic cells.

Life cycle of Actinomycetes

Streptomycetes are the most widely studied and well known genus of the actinomycete
family. Streptomycetes usually inhabit soil and are important decomposers. They also
produce more than half of the world's antibiotics, and are consequently invaluable in the
medical field.

Genome Structure

The entire genome of Streptomyces coelicolor was sequenced as of July 2001. The linear
chromosome is 8,667,507 bp long and is predicted to contain 7,825 genes, about twice as
many as typical free-living bacteria, making it the largest bacterial genome yet
sequenced. The linear chromosome replicates from a central origin. The single
chromosome also has a unique telomere structure. During replication the 5' end of the
chromosome remains incomplete, resulting in a single strand of DNA at each end. The
single strands have several replicating sequences that double over to form hairpin
structures at either end of the chromosome, forming protective telomeres (Goshi et al.).
Streptomyces avermitilis was also been sequenced in October 2001. It is 9,025,608 bp
long, and has 7,575 ORFs assigned. This organism is a well known producer of the anti-
parasitic agent avermectin which is widely used to rid livestock of worm and insect
infestations and to protect large numbers of people from river blindness in sub-Saharan
Africa. There are also two genome projects in the works for Streptomyces scabiei and
Streptomyces ambofaciens.

Cell Structure and Metabolism

The filamentous mycelium of Streptomyces. From: Natural Resources Conservation

Service.

Streptomycetes resemble fungi in their structure. Their branching, filamentous

arrangement of cells form a network called a mycelium. They are able to metabolize
many different compounds including sugars, alcohols, amino acids, and aromatic
compounds by producing extracellular hydrolytic enzymes. Their metabolic diversity is
due to their extremely large genome which has hundreds of transcription factors that
control gene expression, allowing them to respond to specific needs.

Life Cycle
In addition to echoing fungi in their cellular structure, streptomycetes also resemble fungi
in their elaborate life cycle. During the vegetative growth stage of streptomycete
development, DNA replication takes place without cellular division, creating the
previously mentioned filamentous structure. Streptomycetes reproduce and disperse
through the formation of spores, called conidia, which follows the period of vegetative
growth. The spores are produced in aerial filaments called sporophores, which rise above
the colony. Because the complex life cycle of streptomycetes resembles that of
multicellular eukaryotes, it enables researchers to study the development of these more
complex systems using a simpler system.

Ecology

Streptomycetes are found worldwide in soil, and are largely responsible, through the
secretion of chemicals called geosmens, for the earthy smell of soil. streptomycetes
consequently play an important role in the degradation of organic matter, most commonly
noted in compost piles.

Several species of Streptomyces are involved in a symbiotic relationship with species of

ants in the genus Attini. Attine ants cultivate fungus in, what are termed fungal gardens.
They perform all the motions of human farmers, weeding, and nurturing their gardens.
The small bacterium in the streptomyces genus inhabits the cuticles of the ants, and aids
in weeding their fungal gardens. Streptomycetes produce toxins that keep the main weed
in ant fungal gardens, another fungi, Escovopsis, at bay.
Common scab caused by Streptomyces scabies on four potatoes. From: Vegetable MD
Online.

Because streptomycetes inhabit soil, they are mainly phytopathogens, known for
attacking root vegetables, such as potatoes, beets, radishes, rutabaga, turnips, carrots, and
parsnip. Most commonly found on potatoes, Streptomyces scabies creates a condition
known as "common scab," which manifests itself as sores on the external surface of the
potato. The scabs do not harm the meat on the the inside of the potato but create an
extremely unpleasant appearance that devalues the potato.

Phages

Streptomyces coelicolor has a unique bacteriophage resistance system, designed to ward

of the temperate bacteriophage phiC31. The phage growth limitation system of
Streptomyces coelicolor causes phages replicated in a streptomycete cell to become
modified, which activates a mechanism to inhibit phage growth on reinfection of the
same host. Essentially, they manufacture immunity towards bacteriophages.

Medicine

An antibiotic droplet secreted from a Streptomyces colony. From: Higher Education and
Research Opportunities, the John Innes Centre.
The blue haloes surrounding these Streptomyces coelicolor colonies are secreted
actinorhodin, an antibiotic (not yet used clinically). From: The John Innes Centre.

Streptomycetes are most widely known for their ability to synthesize antibiotics. Over 50
different antibiotics have ben isolated from streptomycetes, providing most of the world's
antibiotics. With the newly sequenced genome of Steptomyces coelicolor comes the
possibility of deriving still more antibiotics that have so far remained undiscovered.

The genes of the unusually large genome of Streptomyces coelicolor are grouped together
in clusters, each cluster making a different antibiotic chemical. The genome contains
around 20 clusters, of which only four were previously known. Projects to use the
genome data in the synthesis of new antibiotics are already underway. Many of these
projects are focusing on the possibility to using genetic engineering to to create entirely
new chemicals by splicing together machinery from the numerous templates provided by
Streptomyces coelicolor.

Bleomycin is an antibiotic drug with anticancer properties produced by Streptomyces

verticillus. It was isolated in 1966 by Umezawa et al. and its mechanism of action is
breaking the DNA double helix by the production of free radicals. It is used for malignant
tumors, specifically germ cell tumors, lymphomas, head and neck and Kaposi's sarcomas.

Life cycle of yeast

Budding Yeast: Saccharomyces cerevisiae

Saccharomyces cerevisiae, the

budding yeast, is the common yeast
used in baking ("baker's yeast") and
brewing ("brewer's yeast"). (It is
only distantly related to another
unicellular fungus,
Schizosaccharomyces pombe, the
fission yeast.)

It is a popular "model" organism in

the laboratory because it is a
 unicellular eukaryote whose
cellular activities are much more
like ours than a bacterium like
E. coli. But like E. coli,
 It can be cultured easily.
 It grows rapidly.
 Its entire genome is
known. [Link]
 It can be easily
transformed with genes
from other sources.

Life Cycle

Budding yeast can live with

either two genomes (diploid) or
one (haploid). In either case, it
reproduces by forming buds
(hence the name) by mitosis.

Haploid cells occur in two different mating types: a or α. The type is determined by the
expression of a gene at an active mating type locus.

Haploid cells can live indefinitely in the haploid condition. However, if two cells of
opposite mating types meet, they can fuse and enter the diploid phase of the cell cycle.

This is not as rare event as you might expect.

 Germination of the haploid spores takes place while they are still within the ascus
and mating normally occurs there.
 Even if haploid cells go through a period of growth, they can still find cells of the
opposite mating type most of the time. Although the illustration shows each
haploid cell producing a bud of the same mating type, often the cell switches
mating type. It is able to do so because in addition to the active mating type locus,
it contains two "silent" loci - one a and one α. A double-strand break (DSB) at the
active locus is repaired with the information from one of the silent loci. If the cell
is a, it prefers to tap the information in the silent α locus; and vice-versa.

Cells in the diploid phase are more resistant to harsh environmental conditions. When
diploid cells begin to run out of food, they undergo meiosis, forming four haploid spores
in an ascus (Saccharomyces cerevisiae belongs to the ascomycetes.)

When good conditions return, the spores germinate producing four haploid yeast cells:
two a and two α.
Bacteriophage life cycle

Elsewhere in this museum it is described how infectious diseases can be caused by

viruses. These minute creatures live parasitic in other cells. Some have viruses have
adapted to live in bacterial cells. They are called bacteriophages.

Bacteriophages, viruses that live in bacteria, can have different shapes, and some even
resemble little space-shuttles.

 Have a look at bacteriophages. (Source: Cells Alive)

Bacteriophages are able to attach themselves to (certain types of) bacteria and to inject
their genetic material in the bacterial cell. After this happens, their existence becomes
more difficult to describe, for only their genetic material (in the form of DNA) exists in
the bacterial cell. Then, using the bacterial machinery, the DNA multiplies itself.
Eventually from this multiplied genetic information so many new bacteriophages are
formed that the cell bursts. The offspring of the bacteriophage has destroyed its bacterial
host, and in so doing millions of new bacteriophages are released. These can attach
themselves to new bacteria to complete their life cycle.

How do we notice that bacteriophages are present in bacteria? We can see bacteria when
they grow, eg. a liquid will become cloudy if there are enough bacteria present (check
how our senses can detect bacteria). When we grow bacteria in the laboratory on agar
plates, they can completely cover this plate with a confluent layer of growth. If 
bacteriophages are present in this culture, they will produce holes in this confluent layer.
The hole, called a 'plaque', is circular because phages cannot move, so when the
originally infected bacterial cell bursts, the neighboring cells become infected only. This
process produces a round-shaped spread of phages, thus a perfectly round 'plaque'. A
liquid that is cloudy from bacteria will become clear again when bacteriophages destroy
all cells present. However, bacteriophages do not always kill their host, at least not
immediately. Like all infectious organisms, bacteriophages could never make their host
extinct, for they are dependent on their host bacteria to survive. This is also true for
infectious diseases: they cannot make their target host extinct. See our display on
infectious bacterial diseases.

When bacteriophages kill bacteria, can they be used to cure us from pathogenic bacteria?
Indeed, phage therapy has been used in the past to cure infectious diseases, especially in
Eastern Europe and Russia, and research is continuing. Can phages provide a Nature-
friendly way to fight bacteria? (Source: BI Koerner). BBC Radio provides a transcript of a
program on phage therapy (Source: BBC).

Bacteriophages are important tools in bacteriological research. They can be used as

vehicles to introduce DNA into a bacteria. Phages don't mind to take along foreign DNA
when they insert their own DNA into a bacterial cells. Microbiologists can make use of
this. However, they make sure to keep the phages under control otherwise all their
precious bacterial cultures become infected! The formation of mutations in a bacterial
population can be nicely demonstrated with bacteriophages.

A lytic or lysogenic cycle:

 Lytic, coming from the same stem word for lysis,

means to cut/split. When a bacteriophage infects a
bacterium on a lytic mission, the host cell machinery is
used to produce bacteriophage components. Once a
critical mass is attained, the bacterial cytosol is filled
with bacteriophages and the host cell is sacrificed as it
spills open and releases phages into the environment.
 Lysogenic cycles are much less barbaric. Here, the
bacteriophage integrates itself into the bacterial
genome. When this occurs, the bacteriophage is called
a provirus and it is replicated with each generation of
bacteria. This can occur indefinitely or be triggered to
cause the bacteriophage to enter a lytic cycle.

Why are viruses obligate intracellular parasites?

Viruses do not have the machinery to replicate without a host.

In the case of bacteriophages, the host is a bacteria, e.g.
, and the bacterial ribosomes and machinery are used
to generate the parts to produce progeny viruses.

What are the steps of viral infection (on a cellular level)?

1. Attachment
2. Injection
3. Replication
4. Expression
5. Assembly
6. Release

What makes the life cycle of retroviruses unique?

Retroviruses have an RNA genome that uses a

to turn the genome into DNA (unique

among viruses). The DNA genome then integrates into the
host's genome (common among viruses).