Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Chapter 3 Procaryotic Cell Structure and Function 1 A Preview of Procaryotic Cell Structure and Function • Procaryotes differ from eucaryotes in many traits including size and lack of internal membrane systems • Procaryotes are divided into Bacteria and Archaea • Bacteria are divided into 2 groups based on their Gram stain reaction 2 3 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.2 4 Size, Shape, and Arrangement – Cocci (coccus) – spheres – Diplococci (diplococcus) – pairs – Streptococci – chains – Staphylococci – grape-like clusters – Tetrads – 4 cocci in a square – Sarcinae – cubic configuration of 8 cocci 5 Size, Shape, and Arrangement • Bacilli (bacillus) – rods -coccobacilli – very short rods -vibrios – resemble rods, comma shaped • Spirilla (spirillum) – rigid helices • Spirochetes – flexible helices • Mycelium – network of long, multinucleate filaments 6 Size, Shape, and Arrangement • Pleomorphic – organisms that are variable in shape 7 Size Size range for prokaryotes: 0.2 µm to > 700 µm in diameter – Most cultured rod-shaped bacteria are between 0.5 and 4.0 µm wide and < 15 µm long – Few very large prokaryotes; examples include • Epulopiscium fishelsoni • Thiomargarita namibiensis • Size range for eukaryotic cells: 10 to > 200 µm in diameter 8 9 •largest – 50 μm in diameter • smallest – 0.3 μm in diameter 10 Figure 3.4 11 Procaryotic Cell Membranes • Membranes are an absolute requirement for all living organisms • Plasma membrane encompasses the cytoplasm • Some procaryotes also have internal membrane systems 12 Composition • Phospholipids • Contain both hydrophobic and hydrophilic components • Can exist in many different chemical forms due to variation in the groups attached to the glycerol backbone • Fatty acids point inward to form hydrophobic environment 13 Bacterial Phopholipids • polar ends – interact with water – hydrophilic • nonpolar ends – insoluble in water – hydrophobic 14 Figure 3.6 General Structure of Phospholipids 15 Functions of the plasma membrane • Separation of cell from its environment • Selectively permeable barrier – some molecules are allowed to pass into or out of the cell – transport systems aid in movement of molecules 16 More functions… • Location of crucial metabolic processes • Special receptor molecules in the membrane 17 Fluid Mosaic Model of Membrane Structure • Lipid bilayer in which proteins float Figure 3.5 18 Copyright © The McGraw-Hill Companies, Inc. 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Membrane proteins • Peripheral proteins – loosely associated with the membrane and easily removed • Integral proteins – embedded within the membrane and not easily removed 19 Bacterial Membranes • differ from eucaryotes in lacking sterols – do contain hopanoids, sterol-like molecules • a highly organized, asymmetric system which is also flexible and dynamic 20 Other membrane lipids Figure 3.7 21 Internal Membranous Structures • Plasma membrane infoldings – Observed in many photosynthetic bacteria – Nitrifying bacteria – Methanotrophic bacteria 22 Plasma membrane infoldings: observed in many photosynthetic bacteria Figure 3.8 23 Archaeal membranes • Composed of unique lipids • Some have a monolayer structure instead of a bilayer structure 24 Figure 3.9 25 Figure 3.10 26 Figure 3.11 27 Cytoplasmic Matrix • Substance in which nucleoid, ribosomes and inclusion bodies are suspended • Lacks organelles bound by unit membranes • Composed largely of water 28 Inclusion bodies • Granules of organic or inorganic material that are stockpiled by the cell for future use • Some are enclosed by a singlelayered membrane 29 Organic inclusion bodies • Glycogen – polymer of glucose units • Poly-β-hydroxybutyrate (PHB) – polymers of β-hydroxybutyrate 30 Organic inclusion bodies • cyanophycin granules – large polypeptides containing about equal quantities of arginine and aspartic acid • carboxysomes – contain the enzyme ribulose-1,5,bisphosphate carboxylase (Rubisco), enzyme used for CO2 fixation 31 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.13 32 Organic inclusion bodies • gas vacuoles – found in cyanobacteria and some other aquatic procaryotes – provide buoyancy – aggregates of hollow cylindrical structures called gas vesicles 33 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.14 34 Inorganic inclusion bodies • Polyphosphate granules – linear polymers of phosphates • Sulfur granules - source of energy (purple sulfur bacteria • magnetosomes – contain iron in the form of magnetite – used to orient cells in magnetic fields 35 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. TA 3.2 (a) 36 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. TA 3.2 (b) 37 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. TA 3.2 (c) 38 Ribosomes • Complex structures consisting of protein and RNA • Sites of protein synthesis • Smaller than eucaryotic ribosomes – procaryotic ribosomes 70S – eucaryotic ribosomes 80S • S = Svedburg unit 39 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.15 40 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. The Nucleoid • Irregularly shaped region • Location of chromosome – usually 1/cell • Not membranebound 41 Figure 3.4 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Procaryotic chromosomes are located in the nucleoid, an area in the cytoplasm Figure 3.16 42 The procaryotic chromosome • Closed circular, double-stranded DNA molecule • Looped and coiled extensively • Nucleoid proteins probably aid in folding – nucleoid proteins differ from histones 43 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Plasmids • • • • 44 Closed circular or linear DNA molecules Replicate independently of chromosome Have relatively few genes present Genes on plasmids are not essential to host but may confer selective advantage (e.g., drug resistance) Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Table 3.3 45 The Bacteria Cell Wall • rigid structure that lies just outside the plasma membrane Figure 3.4 46 Functions of cell wall • Provides characteristic shape to cell • Protects the cell from osmotic lysis • Protects from toxic compounds and pathogens • May also contribute to pathogenicity • A site for antibiotic action • Some organisms lack cell walls 47 Cell walls of Bacteria • Bacteria are divided into two major groups based on the response to gramstain procedure. – gram-positive bacteria stain purple – gram-negative bacteria stain pink • Staining reaction due to cell wall structure 48 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.17 49 Peptidoglycan 50 Peptidoglycan (Murein) Structure • Meshlike polymer composed of identical subunits • Contains N-acetyl glucosamine and Nacetylmuramic acid and several different amino acids • Chains of peptidoglycan subunits are cross linked by peptides 51 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.20 52 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.21 53 Gram-Positive Cell Walls • composed primarily of peptidoglycan • may also contain large amounts of teichoic acids • some gram-positive bacteria have layer of proteins on surface of peptidoglycan Figure 3.22 54 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.23 55 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. teichoic acids • polymers of glycerol or ribitol joined by phosphate groups Figure 3.24 56 Gram-Negative Cell Walls • Consist of a thin layer of peptidoglycan surrounded by an outer membrane • Outer membrane composed of lipids, lipoproteins, and lipopolysaccharide (LPS) • No teichoic acids 57 Gram-Negative Cell Walls • More complex than Gram + walls • Peptidoglycan is ~2-5% of wall weight • Periplasmic space differs from that in gram + cells – may constitute 20-40% of cell volume – many enzymes present in periplasm 58 Gram-Negative Cell Walls • Outer membrane lies outside the thin peptidoglycan layer • Braun’s lipoproteins connect outer membrane to peptidoglycan 59 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.25 60 Lipopolysaccharides (LPSs) • Consists of three parts – lipid A – core polysaccharide – O side chain (O antigen) 61 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.27 62 Importance of LPS • Protection from host defenses (O antigen) • Contributes to negative charge on cell surface (core polysaccharide) • Helps stabilize outer membrane structure (lipid A) • Can act as an exotoxin (lipid A) 63 Other Characteristics of the Outer Membrane • More permeable than plasma membrane due to presence of porin proteins and transporter proteins – porin proteins form channels through which small molecules (600-700 daltons) can pass 64 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.28 65 Molecular Model of Porin Proteins 66 Archaeal cell walls • Lack peptidoglycan • Cell wall varies from species to species but usually consists of complex heteropolysaccharides • Methanogens have walls containing pseudomurein 67 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.30 68 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.31 69 Protein Secretion in Procaryotes • Numerous protein secretion pathways have been identified • four major pathways are: – Sec-dependent pathway – type II pathway – type I (ABC) protein secretion pathway – type III protein secretion pathway 70 Overview of Bacterial Protein Secretion • Gram+ and Gram- bacteria have different problems secreting proteins based on the differences between the structure of their walls • Both use Sec-dependent pathway • All protein secretion systems require energy 71 Sec-Dependent Pathway • Also called general secretion pathway • Translocates proteins from cytoplasm across or into plasma membrane • Secreted proteins synthesized as preproteins having amino-terminal signal peptide – signal peptide delays protein folding – chaperone proteins keep preproteins unfolded 72 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Sec-Dependent Pathway • secA translocates preprotein through the plasma membane • When preprotein emerges from plasma membrane a signal peptidase removes the signal peptide 73 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Sec-Dependent Pathway Figure 3.32 74 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Protein Secretion in Grambacteria • 5 secretion pathways have been identified in gram- bacteria • type II and V pathways transport proteins across outer membrane that were translocated across plasma membrane by Sec-dependent pathway • types I and III pathways are Secindependent • type IV pathway usually functions independently of the Sec pathway 75 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Type II Protein Secretion Pathway • Present in a number of plant and animal pathogens • transports proteins from periplasmic across outer membrane • observed in some gram-negative bacteria, including some pathogens • complex systems consisting of up to 12-14 proteins – most are integral membrane proteins 76 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Type V Protein Secretion Pathway • Most recently discovered protein secretion system • Rely on Sec-dependent pathway to move proteins across plasma membrane • When proteins are in periplasmic space many can form a channel in outer membrane through which they transport themselves; hence they are called autotransporters 77 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. ABC Protein Secretion Pathway • also called Type I Protein Secretion Pathway • ubiquitous in procaryotes • transports proteins from cytoplasm across both plasma membrane and outer membrane • secreted proteins have C-terminal secretion signals • Gram+ bacteria use a modified version of this pathway to translocate proteins across the plasma membrane 78 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Type III Protein Secretion Pathway • secretes virulence factors of gramnegative bacteria from cytoplasm, across both plasma membrane and outer membrane, and into host cell • some type III secretion machinery is syringe-shaped – secreted proteins thought to move through a translocation channel 79 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Type IV Protein Secretion Pathway • Type IV pathways are unique because they secrete proteins and transfer DNA during conjugation • Type IV systems are made of many different proteins, some of which form a syringe-like structure 80 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Protein Secretion Systems of Gram-Negative Bacteria Figure 3.33 81 Components External to Cell Wall Figure 3.4 82 Capsules, Slime Layers, and S-Layers Capsules • usually composed of polysaccharides • well organized and not easily removed from cell Slime layers • similar to capsules except diffuse, unorganized and easily removed 83 Bacterial Capsules Figure 3.34 84 Bacterial capsules 85 Glycocalyx – network of polysaccharides extending from the surface of the cell – a capsule or slime layer composed of polysaccharides can also be referred to as a glycocalyx 86 Bacterial Glycocalyx Figure 3.35 87 Capsules, Slime Layers, and SLayers • S-layers – regularly structured layers of protein or glycoprotein – in bacteria the S layer is external to the cell wall – common among Archaea, where they may be the only structure outside the plasma membrane 88 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.36 89 Functions • Protection from host defenses (e.g., phagocytosis) • Protection from harsh environmental conditions (e.g., desiccation) • Attachment to surfaces 90 More functions… • Protection from viral infection or predation by bacteria • Protection from chemicals in environment (e.g., detergents) • Facilitate motility of gliding bacteria • Protection against osmotic stress 91 Pili and Fimbriae • Fimbriae (fimbria) – short, thin, hairlike, proteinaceous appendages • up to 1,000/cell – mediate attachment to surfaces – some (type IV fimbriae) required for twitching motility or gliding motility that occurs in some bacteria • Sex pili (pilus) – similar to fimbriae except longer, thicker, and less numerous (1-10/cell) – required for mating 92 Figure 3.37 93 Flagella and Motility Figure 3.4 94 Patterns of Flagella Distribution • Monotrichous – one flagellum • Polar flagellum – flagellum at end of cell • Amphitrichous – one flagellum at each end of cell • Lophotrichous – cluster of flagella at one or both ends • Peritrichous – spread over entire surface of cell 95 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Flagella Distribution Figure 3.38 96 Flagellar Ultrastructure Figure 3.39 (a) and (b) 97 Flagellar Figure 3.39 (c) 98 Ultrastructure The filament • • • • 99 Extends from cell surface to the tip Hollow, rigid cylinder Composed of the protein flagellin Some procaryotes have a sheath around filament The Hook and Basal Body • hook – links filament to basal body • basal body – series of rings that drive flagellar motor Figure 3.39d 100 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Flagellar Synthesis • an example of self-assembly • complex process involving many genes and gene products • new molecules of flagellin are transported through the hollow filament • growth is from tip, not base 101 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.40 102 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. The Mechanism of Flagellar Movement • flagellum rotates like a propeller – in general, counterclockwise rotation causes forward motion (run) – in general, clockwise rotation disrupts run causing a tumble (twiddle) 103 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.41 104 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.42 105 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Other Types of Motility • spirochetes – exhibit flexing and spinning movements of axial filaments which are composed of periplasmic flagella • gliding motility – cells coast along solid surfaces – no visible motility structure has been identified 106 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Chemotaxis • movement towards a chemical attractant or away from a chemical repellent • concentrations of chemical attractants and chemical repellents detected by chemoreceptors on surfaces of cells 107 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.43 108 Figure 3.44 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Chemotaxis Towards Attractant • in presence of attractant (b) tumbling frequency is reduced and runs in direction of attractant are longer 109 Figure 3.45 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Chemotaxis away from repellent • involves similar but opposite responses 110 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. The Bacterial Endospore • formed by some bacteria • dormant • resistant to numerous environmental conditions – heat – radiation – chemicals – desiccation 111 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.46 112 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.47 113 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. What makes an endospore so resistant? • calcium (complexed with dipicolinic acid) • acid-soluble, DNA-binding proteins • dehydrated core • spore coat • DNA repair enzymes 114 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Sporogenesis • Also called endospore formation or sporulation • normally commences when growth ceases because of lack of nutrients • complex multistage process 115 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Figure 3.49 116 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Transformation of endospore into vegetative cell • germination • complex, multistage process Figure 3.50 117 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. Transformation of dormant spores into active vegetative cells • activation – prepares spores for germination – often results from treatments like heating • germination – – – – spore swelling rupture of absorption of spore coat loss of resistance increased metabolic activity • outgrowth – emergence of vegetative cell 118