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5.TheProkaryotes 1.Bacteria
Topics:
Prokaryotesvs.Eukaryotes
Distinguishingfeaturesofprokaryotes
Bacteria&Achaea
TheGramStain
BasicstructuresofBacterialcells
(Bacteria plural;Bacterium sing.)
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Prokaryotesvs.Eukaryotes
Prokaryote
Eukaryote
Cytoplasm
Cytoplasmic/
Plasma
membrane
Celldivision
Chromosomes
(DNA)
Ribosomes
*Smaller(nano micrometer)
*Unboundnucleoid
Circular chromosome(s)
Binaryfission
NoMembranebound
organelles
70Sribosomes
Rudimentarycytoskeleton
Complexcellwalls
Simpleappendages
*Larger(10100m)
*exceptionsexist
Membranebound
organelles,nucleusand
nucleolus

80Sribosomes
ComplexCytoskeleton
SimplecellwallWhenpresent
Complexappendages
Linear DNAwithhistones
Mitosis
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Mitochondria,
#
Chloroplast

Not80Sinribosomes&chloroplast!
#
onlyinsomemicrobialeukaryotes
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Thereismuchdiversityinprokaryote
cells
ButALLprokaryotessharethefollowingtraits,
Compactgenome nucleoid
70Sribosomes
Tightlycoordinatedcellfunctions
Cytoplasmic(cell,inner,plasma)membrane
Most (notall)havethickcomplexoutercellwalls&cell
envelopes
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The cytoplasmic membrane defines the
existence of a cell
Figure 3.4
Fluidmosaicmembrane
Protectcytoplasm
Phospholipidbilayer
Supportmatrix
Exclusionfunctions
Selectivepermeabilitybarrier
Proteinsarethegatekeepers
Multipleessentialfunctions,e.g.
Respiration
Photosynthesis
Transport/export
Anchorforcellappendages
Sensingandcommunication
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The Gram stain Hans Christian Gram 1884) differentiates
two groups of bacteria
- Gram-positive bacteria retain the crystal violet
stain because of their thicker cell wall
- Gram-negative bacteria do not
Figure 2.21
Purplestaineddiplococcus cellsof
Staphylococcuspneumoniae(0.51.0
m) inasputumsample
GramnegativeProteusmirabilis
(12m)cells
Neutrophilcells
Figure 2.22
SeeCourselink fora
videoontheGram
stainprocedure
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Basicstructureofprokaryotecells:
Thechallengesofbeingaprokaryote
Majorityofprokaryotesexistasfreeliving
organisms
Intheirenvironmentstheyaresubjectto
manystresses
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Predation/
competition/
toxins
Osmoticstress
Nutrientlimitation
Fluctuationsin
Temperature,water
availability,pHetc
(FigfromPrescottFig.3.4)
A simplified model of an average bacterium
/pili
ECPs /
(Plasmids)
ECP=Extracellularpolysaccharides
pg.8081,Fig.3.1showsa
forabeautifulschematicof
anEscherichiacolimodelcell
illustratingthecomplexityof
thecellcytoplasm,nucleoid&
cellwall
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Morphologicaldiversityinbacterialcells
1.TheBudgetversion e.g theTenericutes /Mollicutes (=softskin)
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Components:
Nucleoid DNA
Ribosomes
Cytoplasm
Cytoplasmic/plasma
membrane
lackcellwall;
polymorphicinshape
e.g.genusMycoplasma,
Mycoplasmapneumoniae,
M.genitalium hasperhapsthesmallest
genome(580kbp)ofanyselfreplicatingcell
(E.coli=4,600kbp)
Mollicutes
Advantages &Disadvantages
Compact&Small can
squeezethroughspaces
Doesnotrequirealotof
energyormetabolic
capacitytosurvive
Compactsmallgenome
justtheessentials!
Cannotsustainitself are
oftenparasitesofeukaryote
cells(nutrients,
protection)
Fastidiousinculture lacks
capacitytosynthesizemany
essentialnutrients
Amorphousblob..
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Addingacellwall
Peptidoglycan(alsocalledMurein )
Polymerof2sugars&aminoacids
Sugars:Nacetylglucosamine(NAG)&Nacetylmuramic acid(NAM)
Aminoacid/peptidesareattachedtoNAM
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Formsarigidporoussack/cover(murein
succulus)surroundingthecytoplasmic
membrane:protective,confersshape
SeeFig3.13&
31.4intextbook
enzymesresponsibleforitsbiosynthesismakeexcellent
targetsforantibiotics
Penicillininhibitsthetranspeptidase thatcrosslinksthe
peptides
Vancomycin preventscrossbridgeformationbybinding
totheterminalDAlaDAla dipeptide
Unfortunately,thewidespreaduseofsuch
antibioticsselectsforevolutionofresistantstrains
a.Peptidoglycanisuniquetobacteria
Virulencefactor FragmentsofPGinducefever
duringinfection
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Addingthecellwallcont.
PleasereviewvideoontheGrampositivestain(tobe
discussedlater)
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b)TheBasicGrampositive celloption
Components:
Nucleoid DNA
Ribosomes
Cytoplasm
Cytoplasmic/plasma
membrane
Thick(200 80nm)
cellwall:peptidoglycan
Navarre W W, and Schneewind O
Microbiol. Mol. Biol. Rev. 1999;63:174-
229
Dividing
Streptococcusaureus
cells
BasicGrampositivecellwall
Pros
Strong:thickmurein succulus,
Lipoteichoic acids andTeichoic
acids threadthroughitadding
strength
Protectionagainstosmotic
lysis
Impartshape onthecell
..availableinGramstained
purple
Cons
Mustbeprotected
fromLYSOZYME
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SeeFig3.15oftextbook.NoteS
layersmayormaynotbe
present
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Grampositivebacteria Morphological
diversity
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Staphylococcusaureuscells
coccishaped
Formclusters
Bacilluscereuscells
Rodshaped
Streptococcuspyogenes
Cocci,formchains
http://bioweb.uwlax.edu/
c) The Mycobacterial option
Figure 3.18
Mycobacteria are classified with
Gram-positive bacteria
But stain gram-variable
Stained using the Ziehl-Nielsen acid-
fast stain
Mycobacteria have very complex,
thick and hydrophobic cell
envelopes
- Thick peptidoglycan
- Thick Arabinogalactanlayer
- Mycobacterial outermembrane
-unusual membrane lipids,
Mycolicacids
- Thick capsule of lipids &
glycolipids
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Addingthecellwallcont.
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d)TheBasicGramnegative celloption
Components:
Nucleoid DNA
Ribosomes
Cytoplasm
Cytoplasmic/plasma
membrane
Thin (13sheets)of
peptidoglycan
Outermembrane
(containsLPS)
Welldefined
periplasm
Lipopolysaccharides
(LPS)
peptidoglycan
Cellmembrane
BasicGramnegativecellwall
Pros
Hasbuiltinperiplasmic
compartment
protectsPG
storage,scavenging,
detoxifyingproteins
Abletoprotectitselfagainst
toxicmolecules
OM,periplasmic enzymes
Protectionagainstosmotic
lysis
Variousshapes
..availableinGramstained
pink
Cons
Toxic(lipidAofLPS)
ifnotcorrectly
handled!
Canbeenergetically
expensiveto
maintain
Usuallyhavelarger
genomes genes
forOMcomponents
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Selfstudy:findthecellshapesof,
Helicobacterpylori
Pseudomonasaeruginosa,
Escherichiacoli,
Treponema pallidum
Vibriocholerae
Neisseriameningitidis or Neisseriagonorrhoea
Addedextras/options
(i)Slayer
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Figure 3.17
S- (surface) layer
- 5-25 nm thikc
- An additional protective layer
(bacterial chainmail!)
- Contribute to cell shape, protection
against osmotic lysis, predation
- Crystalline layer of thick subunits
- protein or glycoprotein
- Highly ordered
- Flexible to allow movement and
transport of molecules
TEMofarchaeonThermoproteus
tenax Slayer;subunitsarearranged
inregularhighlyorderedarrays/
patterns
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21
EMoffreezeetchedpreparations
ofbacterialcells(TJBeveridge)
Thinsectionofthecellenvelopeof Bacillusbrevis which
includestheplasmamembrane(thebilayer),apeptidoglycan
layerwhichisthinnerthanusualforGrampositivebacteria,
and(astheuppermoststructure)anSlayer.Bar=100nm. TJ
Beveridge 2006.DOI:10.1111/j.15746976.1997.tb00305.x
TJBeveridge,UoG
Sleyter Uetal.2014.DOI:
10.1111/15746976.12063
Slayer applicationsinbiotechnology
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Subunitsselfassembly
Variousapplicationse.g.
liposomes,nanocapsules
canactasscaffoldortemplatefor
buildingproteinsarrays,super
latticesformolecularelectronics
etc
SlidesfromUweBSleytr etal.2014.FEMSMicrobiologyReviews
24FEB2014DOI:10.1111/15746976.12063
http://onlinelibrary.wiley.com/doi/10.1111/1574
6976.12063/full#fmr12063fig0004
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Addedextras/options
(ii)Capsules
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stained capsule of
Rhizobium trifolii
CulturesofNonmucoid andmucoid
Pseudomonasaeruginosa
http://journal.frontiersin.org/Journa
l/10.3389/fmicb.2011.00162/full
Diagram of the group A streptococcal cell covered with an outer hyaluronic acid capsule and
the group A carbohydrate, consisting of a polymer of rhamnose with N-acetylglucosamine
side chains.
Cunningham M W Clin. Microbiol. Rev. 2000;13:470-511
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Addedextras(sec.3.7)
(iii)Thylakoidsandvesicles
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Figure 3.38
Thylakoids:
OnlyfoundinphotosyntheticGramnegativecells
Arespecializedsystemsofextensivelyfoldedlamellae(sheets)
ofmembranes,packedwithphotosyntheticproteins&
pigments
Maximizephotosyntheticcapacityofcells
marinephototrophProchlorococcus
marinus (TEM).
Halochromatium roseum.
Gasvesicles(seenaswhite
spots) buoyancy
Extras cont..
Carboxysomes
Only in Gram-negative bacteria
Contain enzymes (RuBisco) that fix CO
2
Found in all cyanobacteria as well as
some chemoautotrophs
Have a characteristic polyhedral shape
Storage granules
- Glycogen, Polyhydroxybutyrateetc,
for energy
- Sulfur, for oxidation (Fig 3.39)
Magnetosomes (see Fig 2.44)
- Membrane-embedded crystals of
magnetite, Fe
3
O
4
- Orient the swimming of magnetotactic
bacteria
Figure 3.39
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Flagellum / flagella
Swimming motility
Pili or fimbriae are straight
filaments of pilin protein.
- Used in attachment
Sex pili are used in
conjugation.
Stalks are membrane-
embedded
extensions of the cytoplasm.
- Tips secrete adhesion
factors
called holdfasts
Nanotubes are intercellular
connections that pass material
from one cell to the next.
Nanotubes in Bacillus
subtilis (Fig 3.42)
Pili/Fimbriaeonadividing
Salmonellatyphi cell.
Sexpilus
Flagella:arrows
indicatehooks
andbasalbodies
LetsnotforgetthatProkaryoteshave
simpleCytoskeleton
Prescott Fig. 3.12
Figure shows the Cytoskeleton of Bacillus subtilis (gram-
positive rod-shaped) cell.
Actin-like spiral-shaped cables of MreB
cytoskeleton protein.
- spiral the entire length of the cell
- lies adjacent to the cytoplasmic
(E.coli cell expressing GFP-MreB fusion
protein)
Proteinsofthecytoskeleton:
FtsZ Tubulinhomolog,essentialincell
division
MreB actinhomolog,cellshape
determinant