Bacterial Cell Overview

Lack numerous cellular structural components Functional components required for survival and multiplication Average cell ~ 3,500 structural and functional genes ~ 50% structural proteins and enzymes ~ 50% regulatory functions More complex cell wall structure with multiple layers and additional surface layers Cell structure is dynamic and can change due to environment, age of cells, nutritional status, etc.

Gram-negative idealized cell

Basic structural features

CA - capsule CM - cytoplasmic membrane CS - cytoplasm

F - fimbriae FL - flagellum mRNA - messenger RNA NU - nucleoid OM - outer membrane

P - F pilus PE - periplasm in the periplasmic space PG - peptidoglycan (murein) PL - plasmid

RI - ribosome RNP - RNA polymerase SL - surface layer (S-layer) SM - slimer layer

Cell Envelope
Cell envelope = cytoplasmic membrane + cell wall Acquisition of nutrients Protection from environmental stresses Selectively permeable barrier: 1. uptake nutrients and other essential elements 2. efflux waste products 3. resist osmotic stress 4. maintain cell form E. coli cell envelope

Cytoplasmic Membrane
structure and components
Innermost layer of cell envelope consisting of a phospholipid bilayer embedded with proteins Integral proteins with hydrophobic character in the region that traverses the fatty acid bilayer Hydrophilic proteins or metal ions may attach to hydophilic surfaces Dynamic structure

Cytoplasmic Membrane - Functions

Functions: 1. osmotic barrier 2. transport of nutrients and waste products 3. protein anchoring a. synthesis and export of cell wall components b. secretion of extracellular enzymes and structural proteins 4. generation of ATP (energy source)

Gram stain

Photomicrograph of - Gram-positive (blue-purple, S. aureus) - Gram-negative (pink-red, E. coli)

Types of bacterial cells

Gram-negative cells
Cell wall (CW) is thin, but more complex with an outer membrane (OM). 10-20% of CW is peptidoglycan (PG). Remainder of CW consists of polysaccharides, proteins, & lipids. Cell envelope: cell wall - OM, PG, periplasm cytoplasmic membrane

Gram-negative cell wall

OM located outside of the peptidoglycan OM linked to PG by lipoproteins in periplasm Inner OM - phospholipids and proteins Outer OM - LPS Region between the CM and OM called periplasmic space

Lipopolysaccharide of Gram-negative bacteria

KDO, ketodeoxyoctonate Hep, heptose Glu, glucose Gal, galactose GluNac, N-acetylglucosamine GlcN, glucosamine P, phosphate

Major components: lipid A-core polysaccharide-O-specific polysaccharide Lipid A portion of LPS can be toxic and comprises the endotoxin complex. Long chain LPS (O-antigen side chain) composed of repeating oligosaccharide units; great variation; main somatic antigen of Gram-negatives. LPS is in constant motion.

Gram-negative Periplasm
Dense hydrated gel Contains uncrosslinked PG, periplasmic enzymes, proteins, secreted materials, newly synthesized membrane components Reservoir for pilins, S-layer components, secreted virulence factors Functions: 1. Electron transport 2. Transport of small organic and inorganic molecules 3. Degradation of polymers for import into cell 4. Biosynthesis and degradation of cell wall components for cell division 5. Osmotic stabilization of the cell

Membrane Vesicles (Gram-negative)

Small discharged particles of the OM containing OM proteins, LPS, phospholipids, and periplasmic components. Functions: 1. To insert autolysins into foreign bacteria 2. To export periplasmic virulence factors (toxins, proteases, etc.) to eukaryotic cells 3. To shed and disseminate antigenic materials in membrane blebs

Gram-positive cells

Cell wall - dense layer of peptidoglycan (PG) 60-90% of cell wall is PG PG is closely attached to cell membrane (CM)

Gram-positive cell wall

Lacks both an outer membrane and a periplasmic space Contain teichoic acids which are acidic polysaccharides major antigenic structures lipotechoic acids and PG can induce cytokine production and host inflammation Thick cell walls retain crystal violet-iodine dye

Gram-positive peptidoglycan
Also called the murein layer As much as 90% of the CW; 20-40 layers of PG Composed of two sugar derivatives: N-acetylglucosamine (NAG) N-acetylmuramic acid (NAM) cross-linked & cross-branched to small amino acids and DAP (diaminopimelic acid) Imparts mechanical strength and permeability barrier * NAM and DAP are only present in prokaryotes.* ** PG is target of -lactam antibiotics such as penicillin.**

Acid-fast Bacteria
Mycobacteria Cell wall is thick, like that of Gram-positives CW is ~ 60% lipid with much less PG than Gram-positive bacteria Lipid-rich CW contains unusual glycolipids including mycolic acids and arabinogalactan Protection from macrophage digestion CW components are highly immunogenic Acid-fast staining: Carbolfuchsin dye binds to lipids and resists removal by acid-alcohol mixture Slow growers

Capsules
Third protective covering Polysaccharides covalently linked to phospholipid or lipid A Roles: Protect cells from dessication Adhesion Protect cells from phagocytosis

Pathogenesis: Major virulence factor Resistance to innate host defenses Defend against antibody-mediated defenses Subvert immune system as structures resemble host antigens

Exopolysaccharides - Slime Layers

Less tightly bound than capsule Functions: protects cell against drying helps to trap nutrients near cell binds cells together involved in biofilm co-aggregation and adherence Biofilms protect cells from noxious substances (chlorine or antibiotics), from harsh conditions and dessication predation (protozoa or host cells) Less tightly bound than capsule Dental caries/plaque

S-Layers (Surface Layers)

Crystalline proteinaceous coat on outermost envelope

Composed of single, homogeneous protein or glycoprotein species

Functions: cell adhesion molecular sieves avoidance of host immune response by antigenic variation resistance to complement and phagocytosis

Flagella
Filamentous appendage responsible for cell mobility Permits locomotion toward or away from environmental conditions Chemotaxis - sense and react to chemical gradients Organized differently in different bacteria Important motile pathogenic bacteria: Clostridium E. coli Salmonella Vibrio cholerae Pseudomonas Helicobacter pylori

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Bacterial flagella arrangements

polar, monotrichous

polar, amphitrichous; at each end

lophotrichous; tuft at one or both ends

peritrichous; distributed all over

peritrichous; Legionella SEM

Structure of flagella
Helically wound, hollow filament composed of flagellin subunits a hook complex basel body

Complex basal body mediates insertion and rotation. Functions as rotary motor that uses proton gradient for fuel.

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Flagellar structure and attachment

L ring is embedded in LPS P ring in peptidoglycan S-M ring in CM

Mot proteins = flagellar motor; Motor rotates the filament to propel cell through medium. Fli proteins = motor switch

Flagellar Movement

Forward motion - all flagella rotating CCW in bundle. Tumbling - flagella pushed apart with CW rotation. New forward direction - return to CCW rotation.

Forward motion - single flagellum rotating CCW; pushing. Change direction by reversing flagellar rotation to CW; pulling. New forward motion - return to CCW rotation; pushing.

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Conjugative pili
Filamentous structures encoded by genes on conjugative plasmids Mediate intimate contact between cells and allow genetic exchange Part of Type IV bacterial secretion systems that mediate transfer of DNA or proteins into other cells (bacteria or other cells) Contribute to the virulence of pathogens

Fimbriae (pili)
E. coli

Short, hair-like protein projections Mediate adhesion host cell receptors extracellular matrix proteins other bacteria surfaces - teeth, intestines, rocks, etc. Can have many different types of fimbriae Contribute to resistance to phagocytosis Expression is highly regulated and subject to phase variation

S. typhi

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Cytoplasm

Contains large number of proteins, ribosomes, mRNA, tRNA, DNA Ribosomes - translate mRNA into proteins tRNA - required for recognition of amino acid selection for incorporation into proteins Nucleoid - contains chromosomal DNA, DNA binding proteins, & transcriptional machinery

Nucleoid
Distinct nucleoplasmic region Not membrane bound Very dynamic structure Only sites that are exposed can be transcribed Bacterial genome usually consists of one large circular molecule

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Plasmids
Circular DNA molecules that exist and replicate independent of chromosome. Vary in size.

Some are not essential and can be cured with no change in cell phenotype. Many plasmids encode virulence genes, antibiotic resistance, toxin, antibiotic production, bacteriocins, and metabolic functions. Can promote conjugative transfer of DNA virulence. Antibiotic resistance genes often encoded by transposons which are genetic elements that can be exchanged between different plasmids or the chromosome.

Bacteriophages
Viruses that infect bacteria. Dependent on host for life cycle and for protein synthesis. Virulent phages multiply only through the lytic cycle. Temperate phages multiply their DNA peacefully as a prophage in lysogenic development without lysing the cell. When the prophage form of the bacteriophage exists in chromosome or plasmid, the bacterium is termed a lysogen.

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Bacteriophage infections
Lytic cycle - 5 steps 1. Absorption to the cell via specific receptors (proteins, pili, LPS molecules, etc.) 2. Penetration/Injection insertion of DNA into host

Bacteriophage Cycle
Lytic
3. DNA replication 4. Synthesis of new envelope components 5. Release of progeny phage and lysis of host cell

Lysogenic
3. DNA integration

4. Lysogeny

5. Host lysis may be induced by stresses.

Phages can harbor genes that contribute to virulence of bacterial pathogens, particularly toxins, and contribute to horizontal transfer among bacteria.

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