1 Chapter 4 Prokaryotic & Eukaryotic Cells 2 Prokaryotes VS Eukaryotes General Differences Prokaryotes generally small (0.2 - 2 microns diameter & from 2-8 microns long) no nucleus or other membrane bound organelles one circular chromosome (most) no histone proteins associated with DNA cell wall generally contains peptidoglycan (complex polysaccharide) 3 divide by binary fission Eukaryotes large cells "true nucleus" and other membrane bound organelles multiple linear chromosomes histone proteins always associated with DNA cell wall does not contains peptidoglycan divide by mitosis (complex process) 4 Bacterial Shapes Coccus = sphere shaped Bacillus = rod shaped Coccobacillus = very short rod (egg) shaped Vibrio = comma shaped Spirillium = spiral shaped & rigid Spirochete = spiral shaped & flexible Star-Shaped = example: Stella Rectangular – example Haloarcula 5 Bacterial Arrangements Due to plane of division used when new cells form Chains form when they divide in one plane only (“strep…”) Tetrads form when divisions occur in two planes Cube-like structures form when division is in three planes e.g. Sarciniae Grape-like clusters form with irregular divisions (“staph…”) e.g. Staphylococcus 6 Arrangement: notice the plane of division 7 8 What is the plane of division? How do you get from here? to here? 9 10 11 12 Generalized Structure of Bacterium 13 Structure of a Bacterium (outside first!) Features outside the cell wall MAY include: Glycocalyx (‘sugar halo’) – a protective capsule secreted by the cell. This often has the appearance and texture of a ‘slime coating’ Pili – transfer tubes which allow movement of DNA from one bacterium to another – horizontal evolution Fimbriae – attachment filaments present in many G-organisms. These are used to attach to their target host or tissue. In some cases (mutants) when these are not present the organism is less likely to cause disease since it cannot readily attach to its host 14 Movement Structures (we’re still outside the cell wall) Flagella – long whip-like structures seen in some bacteria when ‘twirled’ or beaten (snakelike) these propel bacteria through their environment Flagellar arrangements: how many & where are they found? Monotrichous – single flagellum located at the pole (end) Lophitrichous –multiple flagella located at one end Amphitrichous –flagella located at both ends Peritrichous – flagella scattered over entire surface 15 16 Attachment of Flagella Basal Body is imbedded in the cell wall and cell membrane (protein complex – two rings in G+ and four in GMain filament is attached to basal body by the hook Clinical note: flagellar proteins can be of use in identifying particular strains of some bacteria e.g. E. coli 0157:H7 (Bad beef!) 17 18 More Movement Structures… Axial Filaments, sometimes called endoflagella (internal flagella) these wrap around some spiral shaped bacteria Contraction of these filaments produces a corkscrew-like motion of these bacteria 19 Movement in Response to Stimuli Chemotaxis – movement toward favorable chemicals such as nutrients (positive chemotaxis) or away from harmful chemicals (negative chemotaxis) chemotaxis follows a chemical gradient – movement toward (or away from) stronger concentration of stimulus Phototaxis – movement toward (or away from) light Attractant – a positive stimulus which attracts bacteria Repellent – a negative stimulus which causes bacteria to flee 20 your book discusses attractant/repellent under chemotaxis, these can also be applied to light Bacterial Cell Wall Composed of a network of repeating sugar molecules and proteins called peptidoglycan (and sometimes other molecules) Two sugars are NAM and NAG – a repeating disaccharide N-acetylglucosamine = NAG N-acetylmuramic acid = NAM Peptides lace through the NAG/NAM bundles 21 22 Gram Positive Cell Wall Details 23 Gram Negative Cell Wall Details Forms channel for transport of useful molecules Useful Endotoxin! in recognition shock & of fever some – blood strainsor gastrointestinal 24 Comparison of G+ and G- cell wall G+ G25 Note large amount of peptidoglycan and no second (outer)layer of plasma membrane Note small amount of peptidoglycan and second (outer)layer of plasma membrane Outer Membrane of GComposed of: Phospholipid bilayer Lipopolysaccharides Lipoproteins Functions: Barrier to some antibiotics barrier to attack from some chemicals (detergents, metals…) Proteins called porins allow some useful materials to enter cell Polysaccharide portion useful for identifying some strains 26 One particular lipid, Lipid-A is an endotoxin causing fever & shock (blood or gastrointestinal) Cell /Plasma Membrane Structure: phospholipid bilayer = two layers of phospholipids Arranged tail to tail with ‘heads’ on the extreme outer & inner edges of the membrane and tails pointing to the inside of the membrane contains embedded proteins This arrangement is known as the Fluid Mosaic Model 27 28 Cell Membrane as a Barrier semi-permeable membrane (permits specific things to pass) small molecules and water pass freely charged molecules and larger ones cannot pass freely must be assisted in some fashion Proteins imbedded in the membrane transport substances across the membrane either as pores or by facilitated diffusion 29 Movement Across the Membrane Some methods for moving across membrane (descriptions to follow on later slides) Diffusion Osmosis Passive Transport Active Transport 30 Diffusion – the movement of particles from region of high concentration to low concentration Tends to continue until evenly distributed throughout the medium 31 Osmosis diffusion of water across a semi-permeable membrane (our cell membrane is a semi-permeable membrane) ok backup, what is diffusion? Movement of molecules from an area of high concentration to an area of low concentration (often proceeds until equilibrium) Solutions relative to cell solute concentration can be either: isotonic: equal solute concentration hypotonic: lower concentration of solutes hypertonic: higher concentration of solutes 32 Osmotic pressure: pressure which causes water movement in or out of a cell 33 34 Passive Transport Large pores in the membrane (formed by proteins) allow some molecules to pass through by simple diffusion This is referred to as passive because the cell has to expend no energy for it to occur Successful passage through a pore often based on charge of molecule size of molecule shape of molecule 35 Facilitated Diffusion Protein carrier molecules provide access across the membrane requires no energy from the cell Substances flow along concentration gradient – that is to say, from high concentration where they are plentiful to low concentration where they are scarce (this provides the energy) 36 Active Transport (requires energy) Protein mediated – proteins act as carrier molecules Molecules that are needed inside the cell can be transported against the concentration gradient Often molecules pass through unchanged to be used in cell as is… ALTERNATIVELY cells can perform: Group Translocation – molecules are chemically altered during trip across the membrane in a way that keeps it inside the cell 37 Cytoplasm The fluid portion of the cell – largely (80%) water, this solution also contains proteins, lipids, ions, carbohydrates and other chemicals 38 ‘Nuclear Area’ The DNA of a bacterium is not contained in a nucleus (i.e. not surrounded by a separate membrane) The bacterial chromosome is a continuous circular loop of double-stranded DNA The chromosome is attached to the inner surface of the cell membrane at a spot called the origin (replication begins here) Occasionally smaller loops of DNA occur separate from the chromosome, these are called plasmids 39 Ribosomes These structures are composed of rRNA and protein They are the ‘protein factories’ of the cell Each ribosome consists of two subunits 30S 50S 40 Inclusions Often prokaryotes store nutrients of other molecules in crystals or droplets in the cytoplasm – Some include: Metachromatic granules – inorganic phosphate stores these often stain red in the presence of Methylene Blue they are typical of Cornybacterium diptheriae Polysaccharide granules – starch or glycogen (energy!) stores stain blue (starch) or red-brown (glycogen) in presence of iodine Lipid Inclusions (fat/oil) – energy store – visible with fat soluble dyes 41 More Types Of Inclusions Sulfur Granules – energy for some – Thiobacillus can utilize sulfur as an energy source Carboxysomes – present in bacteria that can use CO2 and photosynthesis to produce energy. These inclusions are an enzyme that assists their metabolism Gas Vacuoles – ‘bubbles’ these help maintain a particular depth so the organism has access to light & nutrients 42 Still More Inclusions Magnetosomes – magnetic clusters of iron oxide. May provide some protection from hydrogen peroxide (occurs in nature, not just in a brown bottle from the drug store). Novel use for bacterial products! Researchers now grow magnetosome producing bacteria and harvest the magnetite to produce magnetic recording media for tape & computer disks 43 Endospores Resting state formed by some bacteria during environmental stress. Spores are highly resistant to drying & temperature extremes. (can survive boiling water for hours!) Spores can remain dormant for thousands of years! Sporultion – the process of forming spores Germination – spore returning to the vegetative (normal) state 44 45 Endospores and Disease • Dry spores are far more likely to become airborne as ‘dust’. • Thus, when considering virulence (ability to produce disease), dry spores are far more dangerous than wet live cultures. 46 Eukaryotic Cells Organisms with a nucleus AND… Membrane bound organelles Plasma membrane: phospholipid bilayer (as prokaryotic cells) Plasma membrane also contains sterols which may help Prevent lysis (rupture) Membrane also contains surface molecules (proteins & carbohydrates) used in cell-cell recognition. Surface molecules may provide attachment sites for bacteria 47 48 Cell Wall Cell wall: in plants, algae, and fungi (NOT PRESENT in animal cells) Cell wall of yeast = polysaccharides chitin glucan mannan Cell wall of plants (and some algae) also polysaccharide cellulose 49 Cytoplasm Cytoplasm: area inside the plasma membrane and outside nucleus Consists of: cytosol organelles (‘little organs’) Cytosol – the fluid part of the cell (excluding the organelles) about 80% water. Includes dissolved solids such as ions, proteins, other molecules contains filaments (microfilaments, intermediate filaments, and microtubules) which form cytoskeleton 50 Filaments also help accomplish cytoplasmic streaming Cytoplasmic Inclusions ‘Things inside a cell’ Oil/fat droplets – for energy storage Glycogen grains (animal starch) for energy storage Organelles – ‘cellular organs’ 51 Organelles… (so we begin) Ribosomes – the ‘protein factories’ of the cell Made up of rRNA and Protein There are two subunits a large 60S subunit and a small 40S subunit Together then form an 80S ribosome S = refers to their sedimentation rate not their size so you don’t add the numbers 52 Nucleus Nucleus: contain DNA bound to histones and surrounded by double-membrane (nuclear envelope) with small pores (nuclear pores) Nucleoli - condensed regions of DNA where ribosomal RNA is being synthesized 53 54 Mitochondria Production of energy for the cell. Mitochondria have both an inner and outer membrane (double membrane) Synthesize ATP - Enzymes within the matrix (fluid) are involved in the production of ATP Cristae – folds in the inner membrane – site of enzymes involved in ATP production Mitochondria have their own DNA (circular) & ribosome (70S) Mitochondria replicate by binary fission 55 56 Chloroplast Membranous inner sacs called thylakoids contain the chlorophyll and enzymes involved in photosynthesis Chloroplasts have their own DNA & 70S ribosomes Chloroplasts replicate within the cell by binary fission 57 58 Endosymbiotic Theory Chloroplasts & Mitochondria – ‘hitchhikers’ likely to be ancient organisms living in symbiosis with eukaryotic cells They receive safe, nutrient rich environment they provide energy or harvest light for the host cell Both Mitochondria and Chloroplasts: Have circular chromosomes Have 70s Ribosomes Reproduce by binary fision Endosymbiotic Theory 59 Student Reading Review remaining organelles… know BASIC functions of Endoplasmic reticulum Golgi apparatus Lysosomes Peroxisomes Flagella Cilia 60